Anti-CSB antibody [EPR27019-44] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-CSB antibody [EPR27019-44] - BSA and Azide free (AB316745)

This data was developed using ab316744, the same antibody clone in a different buffer formulation.

CSB was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell fresh lysate with ab316744 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316744 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell fresh lysate
Lane 2 : ab316744 IP in HeLa (human cervix adenocarcinoma epithelial cell) whole cell fresh lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab316744 in HeLa whole cell fresh lysate

All lanes:

Immunoprecipitation - Anti-CSB antibody [EPR27019-44] (<a href='/en-us/products/primary-antibodies/csb-antibody-epr27019-44-ab316744'>ab316744</a>) at 1/30 dilution

All lanes:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell fresh lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CSB antibody [EPR27019-44] - BSA and Azide free (AB316745)

This data was developed using ab316744, the same antibody clone in a different buffer formulation.

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-CSB antibody [EPR27019-44] (<a href='/en-us/products/primary-antibodies/csb-antibody-epr27019-44-ab316744'>ab316744</a>) at 1/1000 dilution

Lane 1:

U-2OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 200 kDa,124 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CSB antibody [EPR27019-44] - BSA and Azide free (AB316745)

This data was developed using ab316744, the same antibody clone in a different buffer formulation.

This antibody was tested on fresh and frozen HeLa and 293T Whole Cell Lysate. The Lysates in lanes 3-4 were freshly made and used for Western Blotting immediately to minimize protein degradation.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

Exposure time : Lane 1-2 : 103 seconds; Lane 3-4 : 180 seconds

All lanes:

Western blot - Anti-CSB antibody [EPR27019-44] (<a href='/en-us/products/primary-antibodies/csb-antibody-epr27019-44-ab316744'>ab316744</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

HeLa whole cell lysate at 20 µg

Lane 4:

293T whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 200 kDa,124 kDa

false

• WB

Supplier Data

Western blot - Anti-CSB antibody [EPR27019-44] - BSA and Azide free (AB316745)

This data was developed using ab316744, the same antibody clone in a different buffer formulation.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-CSB antibody [EPR27019-44] (<a href='/en-us/products/primary-antibodies/csb-antibody-epr27019-44-ab316744'>ab316744</a>) at 1/1000 dilution

Lane 1:

Hela (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 40 µg

Lane 2:

HeLa transfected with siRNA specifically targeting ERCC6 whole cell lysate at 40 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 200 kDa,124 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-CSB antibody [EPR27019-44] - BSA and Azide free (AB316745)

This data was developed using the same antibody clone in a different buffer formulation (ab316744).

Western blot : Anti-ERCC6 antibody [EPR27019-44] (ab316744) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab316744 was shown to bind specifically to ERCC6. A band was observed at 180 kDa in wild-type A549 cell lysates with no signal observed at this size in ERCC6 knockout cell line. To generate this image, wild-type and ERCC6 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 fluorescent western blot (TBS-based) blocking solution 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-CSB antibody [EPR27019-44] (<a href='/en-us/products/primary-antibodies/csb-antibody-epr27019-44-ab316744'>ab316744</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

ERCC6 knockout A549 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

PC-3 Cytoplasmic Fraction 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: 180 kDa

false