Anti-GAPDH antibody [6C5] - Loading Control

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

ab8245 staining GAPDH in SV40LT-SMC (Rat SV40-transfected aorta smooth cell line) cells.

The cells were fixed with 4% formaldehyde (10 minutes) permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab8245 at 5μg/ml and ab202272 at 1/250 dilution overnight at +4°C followed by a further incubation at room temperature for 1h with Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150117) (shown in green). Nuclear DNA was labeled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

ab8245 staining GAPDH in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

The cells were fixed with 100% methanol (5 minutes) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab8245 at 5 μg/ml and ab6046 at 1 μg/ml overnight at +4°C followed by a further incubation at room temperature for 1 hour with Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150117) at 2 μg/ml (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) preadsorbed (ab150088) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labeled in blue with DAPI.

Negative controls : 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

ab8245 staining GAPDH in NIH/3T3 (Mouse embryo fibroblast cell line) cells.

The cells were fixed with 4% formaldehyde (10 minutes) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab8245 at 1 μg/ml and ab202272 at 1/250 dilution overnight at +4°C followed by a further incubation at room temperature for 1 hour with Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150117) (shown in green). Nuclear DNA was labeled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-SSB antibody [EPR6570] ab124932 staining at 1/10000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 49 kDa in Wild-type HeLa cell lysates with no signal observed at this size in SSB knockout HeLa cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-SSB antibody [EPR6570] (<a href='/ko/products/primary-antibodies/ssb-antibody-epr6570-ab124932'>ab124932</a>) at 1/10000 dilution

Lane 1:

Wild-type HeLa cell lysates at 20 µg

Lane 2:

Western blot - Human SSB knockout HeLa cell line (<a href='/ko/products/cell-lines/human-ssb-knockout-hela-cell-line-ab274949'>ab274949</a>) at 20 µg

Lane 3:

HCT 116 cell lysates at 20 µg

Lane 4:

PC-3 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 47 kDa

Observed band size: 49 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

All lanes:

Western blot - Anti-Chk1 antibody [E250] (<a href='/ko/products/primary-antibodies/chk1-antibody-e250-ab32531'>ab32531</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

CHEK1 knockout A549 cell lysate at 20 µg

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

MOLT-4 cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 54 kDa

Observed band size: 57 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-PRKCD antibody [EPR17075] (ab182126) staining at 1/5000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab182126 was shown to bind specifically to PRKCD. A band was observed at 75-80 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in PRKCD knockout cell line. To generate this image, wild-type and PRKCD knockout U-87 MG cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-PKC delta antibody [EPR17075] (<a href='/ko/products/primary-antibodies/pkc-delta-antibody-epr17075-ab182126'>ab182126</a>) at 1/5000 dilution

Lane 1:

Wild-type U-87 MG cell lysate at 20 µg

Lane 2:

PRKCD knockout U-87 MG cell lysate at 20 µg

Secondary

Lanes 1 - 2:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 2:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 75-80 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-STK11 antibody [EPR19379] (ab199970) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab199970 was shown to bind specifically to STK11. A band was observed at 50-60 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in STK11 knockout cell line. To generate this image, wild-type and STK11 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-LKB1 antibody [EPR19379] (<a href='/ko/products/primary-antibodies/lkb1-antibody-epr19379-ab199970'>ab199970</a>) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

STK11 knockout HCT 116 cell lysate at 20 µg

Lane 3:

Wild-type HEK-293T ab255553 cell lysate at 20 µg

Lane 4:

STK11 knockout HEK-293T ab261047 cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 50-60 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-BACE1 antibody [EPR3956] (ab108394) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab108394 was shown to bind specifically to BACE1. A band was observed at 56 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in BACE1 knockout cell line. To generate this image, wild-type and BACE1 knockout U-87 MG cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-BACE1 antibody [EPR3956] (<a href='/ko/products/primary-antibodies/bace1-antibody-epr3956-ab108394'>ab108394</a>) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG cell lysate at 20 µg

Lane 2:

BACE1 knockout U-87 MG cell lysate at 20 µg

Lane 3:

Wild-type SH-SY5Y cell lysate at 20 µg

Lane 4:

BACE1 knockout SH-SY5Y <a href='/ko/products/unavailable/human-bace1-knockout-sh-sy5y-cell-line-ab280078'>ab280078</a> cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 56 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-TAF15 antibody [EPR9197(B)] ab134916 staining at 1/10000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 74 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in TAF15 knockout U-87 MG cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-TAF15 antibody [EPR9197(B)] (<a href='/ko/products/primary-antibodies/taf15-antibody-epr9197b-ab134916'>ab134916</a>) at 1/10000 dilution

Lane 1:

Wild-type U-87 MG whole cell lysate at 20 µg

Lane 2:

TAF15 knockout U-87 MG whole cell lysate at 20 µg

Lane 3:

Raji whole cell lysate at 20 µg

Lane 4:

HT1080 whole cell lysate at 20 µg

Lane 5:

PC-3 whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 62 kDa

Observed band size: 74 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-KEAP1 antibody staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, anti-KEAP1 antibody was shown to bind specifically to KEAP1. A band was observed at 70 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in KEAP1 knockout cell line. To generate this image, wild-type and KEAP1 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Anti-KEAP1 antibody at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

KEAP1 knockout A549 cell lysate at 20 µg

Lane 3:

Wild-type HCT 116 cell lysate at 20 µg

Lane 4:

KEAP1 knockout HCT 116 <a href='/ko/products/cell-lines/human-keap1-knockout-hct116-cell-line-ab286484'>ab286484</a> cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 70 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-TAB2 antibody [EPR19670-161] ab317602 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 80 kDa in Wild-type A549 cell lysates with no signal observed at this size in TAB2 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-TAB2 antibody [EPR19670-161] (<a href='/ko/products/primary-antibodies/tab2-antibody-epr19670-161-ab317602'>ab317602</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 whole cell lysate at 20 µg

Lane 2:

Western blot - Human TAB2 knockout A549 cell line (<a href='/ko/products/cell-lines/human-tab2-knockout-a549-cell-line-ab301263'>ab301263</a>) at 20 µg

Lane 3:

HEK-293 whole cell lysate at 20 µg

Lane 4:

Raji whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 76 kDa

Observed band size: 80 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-B7-H6 antibody [EPR23470-210] ab253180 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 70-80 kDa in Wild-type Raji UNBOILED cell lysates with no signal observed at this size in NCR3LG1 knockout Raji UNBOILED cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-B7-H6 antibody [EPR23470-210] (<a href='/ko/products/primary-antibodies/b7-h6-antibody-epr23470-210-ab253180'>ab253180</a>) at 1/1000 dilution

Lane 1:

Wild-type Raji UNBOILED whole cell lysate at 20 µg

Lane 2:

Western blot - Human NCR3LG1 knockout Raji cell line (<a href='/ko/products/cell-lines/human-ncr3lg1-knockout-raji-cell-line-ab290414'>ab290414</a>) at 20 µg

Lane 3:

HepG2 UNBOILED whole cell lysate at 20 µg

Lane 4:

K562 UNBOILED whole cell lysate at 20 µg

Lane 5:

Daudi UNBOILED whole cell lysate at 20 µg

Lane 6:

SK-OV-3 UNBOILED whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 51 kDa

Observed band size: 70-80 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-B7-H6 antibody [EPR21841] ab219205 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 70-80 kDa in Wild-type Raji UNBOILED cell lysates with no signal observed at this size in NCR3LG1 knockout Raji UNBOILED cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-B7-H6 antibody [EPR21841] (<a href='/ko/products/primary-antibodies/b7-h6-antibody-epr21841-ab219205'>ab219205</a>) at 1/1000 dilution

Lane 1:

Wild-type Raji UNBOILED whole cell lysate at 20 µg

Lane 2:

Western blot - Human NCR3LG1 knockout Raji cell line (<a href='/ko/products/cell-lines/human-ncr3lg1-knockout-raji-cell-line-ab290414'>ab290414</a>) at 20 µg

Lane 3:

HepG2 UNBOILED whole cell lysate at 20 µg

Lane 4:

K562 UNBOILED whole cell lysate at 20 µg

Lane 5:

Daudi UNBOILED whole cell lysate at 20 µg

Lane 6:

SK-OV-3 UNBOILED whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 51 kDa

Observed band size: 70-80 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-Optineurin Antibody [RP23040250] ab313418 staining at 1 µg/mL, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 73 kDa in Wild-type MCF7 cell lysates with no signal observed at this size in OPTN knockout MCF7 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Optineurin antibody [RP23040250] (<a href='/ko/products/primary-antibodies/optineurin-antibody-rp23040250-ab313418'>ab313418</a>) at 1 µg/mL

Lane 1:

Wild-type MCF7 whole cell lysate at 20 µg

Lane 2:

OPTN knockout MCF7 whole cell lysate at 20 µg

Lane 3:

U-2 OS whole cell lysate at 20 µg

Lane 4:

Raji whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 66 kDa

Observed band size: 73 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-Optineurin antibody [EPR23059-124] ab242146 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 73 kDa in Wild-type MCF7 cell lysates with no signal observed at this size in OPTN knockout MCF7 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Optineurin antibody [EPR23059-124] (<a href='/ko/products/primary-antibodies/optineurin-antibody-epr23059-124-ab242146'>ab242146</a>) at 1/1000 dilution

Lane 1:

Wild-type MCF7 whole cell lysate at 20 µg

Lane 2:

OPTN knockout MCF7 whole cell lysate at 20 µg

Lane 3:

U-2 OS whole cell lysate at 20 µg

Lane 4:

Raji whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 66 kDa

Observed band size: 73 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Lane 1 : Wild-type HCT 116 cell lysate 20 μg
Lane 2 : CTNNB1 knockout HCT 116 cell lysate 20 μg
False colour image of Western blot : Anti-beta Catenin antibody [IGX4794R-3] staining at 1 μg/ml, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab223075 was shown to bind specifically to beta Catenin. A band was observed at 95 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in CTNNB1 knockout cell line ab273712 (knockout cell lysate ab275247). The band observed in the knockout lysate lane below 95 kDa is likely to represent a truncated form of beta Catenin. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and CTNNB1 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-beta Catenin antibody [IGX4794R-3] (<a href='/ko/products/primary-antibodies/beta-catenin-antibody-igx4794r-3-ab223075'>ab223075</a>) at 1 µg/mL

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

Western blot - Human CTNNB1 (beta Catenin) knockout HCT116 cell lysate (<a href='/ko/products/cell-lysates/human-ctnnb1-beta-catenin-knockout-hct116-cell-lysate-ab275247'>ab275247</a>) at 20 µg

Secondary

Lanes 1 - 2:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Lanes 1 - 2:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 85 kDa

Observed band size: 95 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

All lanes:

Western blot - Anti-Presenilin 2/AD5 antibody [EP1515Y] (<a href='/ko/products/primary-antibodies/presenilin-2-ad5-antibody-ep1515y-ab51249'>ab51249</a>) at 1/20000 dilution

Lane 1:

Wild-type U-87 MG ab278079 at 20 µg

Lane 2:

PSEN2 knockout U-87 MG at 20 µg

Lane 3:

A549 at 20 µg

Lane 4:

MOLT-4 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 50 kDa

Observed band size: 18 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-TAF15 antibody [EPR9196(B)] ab133760 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 74 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in TAF15 knockout U-87 MG cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-TAF15 antibody [EPR9196(B)] (<a href='/ko/products/primary-antibodies/taf15-antibody-epr9196b-ab133760'>ab133760</a>) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG whole cell lysate at 20 µg

Lane 2:

TAF15 knockout U-87 MG whole cell lysate at 20 µg

Lane 3:

Raji whole cell lysate at 20 µg

Lane 4:

HT1080 whole cell lysate at 20 µg

Lane 5:

PC-3 whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 62 kDa

Observed band size: 74 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-UNC5B antibody [EPR26173-186] ab313565 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 120, 50 kDa in Wild-type U-87 MG cell lysates with a truncated product observed at 100 kDa in CRISPR-Cas9 Edited cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-UNC5B antibody [EPR26173-186] (<a href='/ko/products/primary-antibodies/unc5b-antibody-epr26173-186-ab313565'>ab313565</a>) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG (UNBOILED) whole cell lysate at 20 µg

Lane 2:

UNC5B knockout U-87 MG (UNBOILED) whole cell lysate at 20 µg

Lane 3:

SH-SY5Y (UNBOILED) whole cell lysate at 20 µg

Lane 4:

U-2 OS (UNBOILED) whole cell lysate at 20 µg

Lane 5:

Daudi (UNBOILED) whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 104 kDa

Observed band size: 120 kDa,50 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-TUG antibody [EPR8615] ab131237 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 70 kDa in Wild-type A549 cell lysates with no signal observed at this size in ASPSCR1 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-TUG antibody [EPR8615] (<a href='/ko/products/primary-antibodies/tug-antibody-epr8615-ab131237'>ab131237</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 whole cell lysate at 20 µg

Lane 2:

ASPSCR1 knockout A549 whole cell lysate at 20 µg

Lane 3:

HEK-293 whole cell lysate at 20 µg

Lane 4:

HeLa whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 70 kDa

Observed band size: 70 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-TUG antibody [EPR8616] ab131217 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 70 kDa in Wild-type A549 cell lysates with no signal observed at this size in ASPSCR1 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-TUG antibody [EPR8616] (<a href='/ko/products/primary-antibodies/tug-antibody-epr8616-ab131217'>ab131217</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 whole cell lysate at 20 µg

Lane 2:

ASPSCR1 knockout A549 whole cell lysate at 20 µg

Lane 3:

HEK-293 whole cell lysate at 20 µg

Lane 4:

HeLa whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 70 kDa

Observed band size: 70 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-RIP3 antibody [EPR24374-135] ab305054 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 57 kDa in Wild-type THP-1 cell lysates with no signal observed at this size in RIPK3 knockout THP-1 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-RIP3 antibody [EPR24374-135] (<a href='/ko/products/primary-antibodies/rip3-antibody-epr24374-135-ab305054'>ab305054</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 whole cell lysate at 20 µg

Lane 2:

Western blot - Human RIPK3 knockout THP-1 cell line (<a href='/ko/products/cell-lines/human-ripk3-knockout-thp-1-cell-line-ab280065'>ab280065</a>) at 20 µg

Lane 3:

Jurkat whole cell lysate at 20 µg

Lane 4:

HeLa whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 57 kDa

Observed band size: 57 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-STAT1 (phospho S727) antibody [EPR3146] ab109461 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 91 kDa. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-STAT1 (phospho S727) antibody [EPR3146] (<a href='/ko/products/primary-antibodies/stat1-phospho-s727-antibody-epr3146-ab109461'>ab109461</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa IFN-γ (100 ng/mL, 30 min) whole cell lysate at 20 µg

Lane 2:

Wild-type HeLa Vehicle control IFN-γ (0 ng/mL, 30 min) whole cell lysate at 20 µg

Lane 3:

STAT1 knockout HeLa IFN-γ (100 ng/mL, 30 min) whole cell lysate at 20 µg

Lane 4:

STAT1 knockout HeLa Vehicle control IFN-γ (0 ng/mL, 30 min) whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 91 kDa

Observed band size: 91 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Rabbit Monoclonal [EPR18513] to PKC ab179524 staining at 1/2000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 76, 85 kDa in Wild-type A549 cell lysates with no signal observed at this size in PRKCH knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-PKC eta antibody [EPR18513] (<a href='/ko/products/primary-antibodies/pkc-eta-antibody-epr18513-ab179524'>ab179524</a>) at 1/2000 dilution

Lane 1:

Wild-type A549 whole cell lysate at 20 µg

Lane 2:

PRKCH knockout A549 whole cell lysate at 20 µg

Lane 3:

U-2 OS whole cell lysate at 20 µg

Lane 4:

HL-60 whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 78 kDa

Observed band size: 76 kDa,85 kDa

false

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

False colour image of Western blot : Anti-MEF2A + MEF2C antibody [EPR19089-34] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab197070 was shown to bind specifically to MEF2C. A band was observed at 55/60 kDa in wild-type THP-1 cell lysates with no signal observed at this size in MEF2C knockout cell line (ab313739). To generate this image, wild-type and MEF2C knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-MEF2A + MEF2C antibody [EPR19089-34] (<a href='/ko/products/primary-antibodies/mef2a-mef2c-antibody-epr19089-34-ab197070'>ab197070</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

MEF2C knockout THP-1 cell lysate at 20 µg

Lane 3:

Daudi cell lysate at 20 µg

Lane 4:

Jurkat cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 55/60 kDa,37 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Anti-LIFR antibody [EPR24651-109] (ab283651) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab283651 was shown to bind specifically to LIFR. A band was observed at 100-175 kDa in wild-type HeLa cell lysates with no signal observed at this size in LIFR knockout cell line (knockout cell lysate ab282984). To generate this image, wild-type and LIFR knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-LIFR antibody [EPR24651-109] (<a href='/ko/products/primary-antibodies/lifr-antibody-epr24651-109-ab283651'>ab283651</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

LIFR knockout HeLa cell lysate at 20 µg

Lane 3:

PC-3 cell lysate at 20 µg

Lane 4:

T-47D cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 124 kDa

false

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

False colour image of Western blot : Anti-PAK2 antibody [EP796Y] staining at 1/5000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab76293 was shown to bind specifically to PAK2. A band was observed at 65 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in PAK2 CRISPR-Cas9 edited cell line ab282648 (CRISPR-Cas9 edited cell lysate ab283047). The band observed in the CRISPR-Cas9 edited lysate lane below 65 kDa is likely to represent a truncated form of PAK2. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and PAK2 CRISPR-Cas9 edited HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-PAK2 antibody [EP796Y] (<a href='/ko/products/primary-antibodies/pak2-antibody-ep796y-ab76293'>ab76293</a>) at 1/5000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human PAK2 knockout HEK-293T cell lysate (ab283047) at 20 µg

Lane 3:

Wild-type HeLa ab255552 cell lysate at 20 µg

Lane 4:

PAK2 knockout HeLa ab260287 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 58 kDa

Observed band size: 65 kDa

false

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

False colour image of Western blot : Anti-FURIN antibody [E1W4O] (ab70393S) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab70393S was shown to bind specifically to FURIN. A band was observed at 80-90 kDa in wild-type Calu-3 cell lysates with no signal observed at this size in FURIN knockout cell line ab275417 (knockout cell lysate ab275533). To generate this image, wild-type and FURIN knockout Calu-3 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Anti-FURIN antibody at 1/1000 dilution

Lane 1:

Wild-type Calu-3 cell lysate at 20 µg

Lane 2:

Western blot - Human FURIN knockout Calu-3 cell lysate (ab275533) at 20 µg

Lanes 3 - 4:

Ramos cell lysate at 20 µg

Lane 5:

U-87 MG cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 80-90 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-CDKN1A antibody [EPR362] (ab109520) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab109520 was shown to bind specifically to CDKN1A. A band was observed at 21 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in CDKN1A knockout cell line. To generate this image, wild-type and CDKN1A knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-p21 antibody [EPR362] (<a href='/ko/products/primary-antibodies/p21-antibody-epr362-ab109520'>ab109520</a>) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

CDKN1A knockout HCT 116 cell lysate at 20 µg

Lane 3:

Wild-type MCF7 cell lysate at 20 µg

Lane 4:

CDKN1A knockout MCF7 cell lysate at 20 µg

Lane 5:

Wild-type A549 cell lysate at 20 µg

Lane 6:

CDKN1A knockout A549 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-RAF1 antibody [EP4969] (ab181115) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab181115 was shown to bind specifically to RAF1. A band was observed at 73 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in RAF1 knockout cell line. To generate this image, wild-type and RAF1 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Raf1 antibody [EP4969] - N-terminal (<a href='/ko/products/primary-antibodies/raf1-antibody-ep4969-n-terminal-ab181115'>ab181115</a>) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

RAF1 knockout HCT 116 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 73 kDa

Observed band size: 73 kDa

false

• WB

Supplier Data

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

False colour image of Western blot : Anti-RHEB antibody staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, the antibody was shown to bind specifically to RHEB . A band was observed at 18 kDa in wild-type C2C12 cell lysates with no signal observed at this size in RHEB knockout cell line ab284606. To generate this image, wild-type and RHEB knockout C2C12 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Anti-RHEB antibody at 1/1000 dilution

Lane 1:

Wild-type C2C12 cell lysate at 20 µg

Lane 2:

RHEB knockout C2C12 cell lysate at 20 µg

Lane 3:

Raji cell lysate at 20 µg

Lane 4:

U-2 OS cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 20 kDa

Observed band size: 18 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-Optineurin antibody [EPR20654] ab213556 staining at 1/500 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 73 kDa in Wild-type MCF7 cell lysates with no signal observed at this size in OPTN knockout MCF7 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Optineurin antibody [EPR20654] (<a href='/ko/products/primary-antibodies/optineurin-antibody-epr20654-ab213556'>ab213556</a>) at 1/500 dilution

Lane 1:

Wild-type MCF7 whole cell lysate at 20 µg

Lane 2:

OPTN knockout MCF7 whole cell lysate at 20 µg

Lane 3:

U-2 OS whole cell lysate at 20 µg

Lane 4:

Raji whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 66 kDa

Observed band size: 73 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-BRD3 antibody [EPR23743-226] (ab300106) staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab300106 was shown to bind specifically to BRD3.

A band was observed at 90-100 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in BRD3 knockout cell line.

To generate this image, wild-type and BRD3 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.

Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-BRD3 antibody [EPR23743-226] (<a href='/ko/products/primary-antibodies/brd3-antibody-epr23743-226-ab300106'>ab300106</a>) at 1/500 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

BRD3 knockout HCT 116 cell lysate at 20 µg

Lane 3:

Wild-type HEK-293T ab255553 cell lysate at 20 µg

Lane 4:

BRD3 knockout HEK-293T ab260529 cell lysate at 20 µg

Secondary

Lanes 1 - 4:

HRP conjugated Goat anti-Rabbit (H+L) at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 90-100 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Blocking buffer and concentration : ½ volume of Odyssey Blocking Buffer (TBS)+ ½ volume of 0.1% TBS                            Diluting buffer and concentration : ½ volume of Odyssey Blocking Buffer (TBS)+ ½ volume of 0.1% TBST    False colour image of Western blot : Anti-Coronin-1C antibody [EPR25365-24] (ab283693) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab283693 was shown to bind specifically to Coronin-1C. A band was observed at 53 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in Coronin-1C knockout cell line ab266381 (knockout cell lysate (ab258377). To generate this image, wild-type and Coronin-1C knockout HEK-293T cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

Lanes 1 - 2:

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) at 1/20000 dilution

Lanes 1 - 2:

Western blot - Anti-Coronin-1C antibody [EPR25365-24] (<a href='/ko/products/primary-antibodies/coronin-1c-antibody-epr25365-24-ab283693'>ab283693</a>) at 1/1000 dilution

Lane 1:

Wild-type 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

Western blot - Human CORO1C (Coronin-1C) knockout HEK-293T cell lysate (<a href='/ko/products/cell-lysates/human-coro1c-coronin-1c-knockout-hek-293t-cell-lysate-ab258377'>ab258377</a>) at 20 µg

Secondary

Lanes 1 - 2:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Lanes 1 - 2:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 53 kDa

Observed band size: 53 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

False colour image of Western blot : Anti-Smad3 antibody [EP568Y] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab40854 was shown to bind specifically to Smad3. A band was observed at 50 kDa in wild-type A549 cell lysates with no signal observed at this size in SMAD3 CRISPR-Cas9 edited cell line ab277888 (CRISPR-Cas9 edited cell lysate None). The band observed in the CRISPR-Cas9 edited lysate lane below 50 kDa is likely to represent a truncated form of Smad3. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and SMAD3 CRISPR-Cas9 edited A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Smad3 antibody [EP568Y] (<a href='/ko/products/primary-antibodies/smad3-antibody-ep568y-ab40854'>ab40854</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

SMAD3 CRISPR-Cas9 edited A549 cell lysate at 20 µg

Secondary

Lanes 1 - 2:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 2:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 48 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

All lanes:

Akt3 (E1Z3W) Rabbit mAb at 1/1000 dilution

Lane 1:

Wild-type A549 ab288558 at 20 µg

Lane 3:

THP-1 at 20 µg

Lane 4:

A549 Cytoplasmic at 20 µg

Lane 5:

Recombinant Human AKT3 protein at 0.2 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 54 kDa

Observed band size: 60 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Rabbit Monoclonal [EP854(2)Y] to gamma H2A.X (phospho S139) ab81299 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% BSA in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-gamma H2A.X (phospho S139) antibody [EP854(2)Y] (<a href='/ko/products/primary-antibodies/gamma-h2ax-phospho-s139-antibody-ep8542y-ab81299'>ab81299</a>) at 1/1000 dilution

Lane 1:

HepG2 Vehicle Control Etoposide (0 uM, 90 min) at 20 µg

Lane 2:

HepG2 Etoposide treated (100 uM, 90 min) at 20 µg

Lane 3:

Jurkat Vehicle Control Etoposide (0 uM, 90 min) at 20 µg

Lane 4:

Jurkat Treated Etoposide (100 uM, 90 min) at 20 µg

Lane 5:

Human Brain at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 15 kDa

Observed band size: 15 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Anti-Iba1 antibody [EPR16588] - Microglia marker - Goat IgG (Chimeric) (ab289874) at 1/1000, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 17 kDa. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Donkey anti-Goat IgG 800CW and Donkey anti-Mouse IgG 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Iba1 antibody [EPR16588] - Microglia marker - Goat IgG (Chimeric) (<a href='/ko/products/primary-antibodies/iba1-antibody-epr16588-microglia-marker-goat-igg-chimeric-ab289874'>ab289874</a>) at 1/1000 dilution

Lane 1:

THP-1 (Human monocytic cell line) cell lysate

Lane 2:

Human spleen tissue lysate

Lane 3:

Human brain tissue lysate

Lane 4:

MCF7 (human breast cancer cell line) cell lysate

Lane 5:

RAW264.7 (mouse macrophage cell line) cell lysate

Lane 6:

Mouse spleen tissue lysate

Lane 7:

Mouse brain tissue lysate

Lane 8:

NIH/3T3 (mouse embryonic fibroblast cell line) cell lysate

Lane 9:

Rat spleen tissue lysate

Lane 10:

Rat brain tissue lysate

Lane 11:

NR8383 (rat alveolar macrophage cell line) cell lysate

Lane 12:

C6 (rat glioma cell line) cell lysate

Secondary

All lanes:

Donkey anti-Goat IgG 800CW and Donkey anti-Mouse IgG 680RD at 1/20000 dilution

Observed band size: 17 kDa,36 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Western blot : Rabbit monoclonal [EPR1216] to SDF1 ab155090 staining at 1/500 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 25 kDa in E2 treated wild-type cell lysates with no signal observed at this size in E2 treated CXCL12 knockout MCF7.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.

Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-SDF1 antibody [EPR1216] (<a href='/ko/products/primary-antibodies/sdf1-antibody-epr1216-ab155090'>ab155090</a>) at 1/500 dilution

Lane 1:

Untreated Wild-type MCF7 cell lysates at 20 µg

Lane 2:

E2 treated wild-type MCF7 cell lysates at 20 µg

Lane 3:

Untreated CXCL12 knockout MCF7 cell lysate at 20 µg

Lanes 3 - 4:

Western blot - Human CXCL12 knockout MCF7 cell line (ab281609)

Lane 4:

E2 treated CXCL12 knockout MCF7 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 25 kDa

Observed band size: 25 kDa,36 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Blocking and diluting buffer and concentration : 3% NFDM/TBST.

Western blot : Rabbit monoclonal [EPR30452-24-2] to NPM3 ab327418 staining at 1 µg/mL, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 23 kDa. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG 800CW and Goat anti-Mouse IgG 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-NPM3 antibody [EPR30452-24-2] (<a href='/ko/products/primary-antibodies/npm3-antibody-epr30452-24-2-ab327418'>ab327418</a>) at 1 µg/mL

Lane 1:

Western blot - Human wild-type HEK-293T cell line (ab255449)

Lane 2:

Western blot - Human NPM3 knockout HEK-293T cell line (<a href='/ko/products/cell-lines/human-npm3-knockout-hek-293t-cell-line-ab266751'>ab266751</a>)

Lane 3:

HeLa

Lane 4:

Mouse Testis

Lane 5:

Mouse Thymus

Lane 6:

Mouse Skeletal Muscle

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (800CW) at 1/20000 dilution

Observed band size: 23 kDa,37 kDa

false

• WB

Lab

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (AB8245)

Anti-CHEK1 antibody [EP691Y] (ab40866) staining at 1/10000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab40866 was shown to bind specifically to CHEK1. A band was observed at 57 kDa in wild-type A549 cell lysates with a reduction in signal observed at this size in CHEK1 heterozygous knockout cell line. To generate this image, wild-type and CHEK1 heterozygous knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Chk1 antibody [EP691Y] (<a href='/ko/products/primary-antibodies/chk1-antibody-ep691y-ab40866'>ab40866</a>) at 1/10000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

CHEK1 knockout A549 cell lysate at 20 µg

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

MOLT-4 cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 54 kDa

Observed band size: 57 kDa

false