Goat Anti-Rat IgG H&L (HRP)

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking and diluting buffer : 5% NFDM/TBST.

The molecular weight is consistent with the benchmark : ab186733.

The positive cells were the consistent with the literature provided by licensor (PMID : 1843089).

All lanes:

Western blot - Anti-Argonaute-2 antibody [11A9] (<a href='/en-us/products/primary-antibodies/argonaute-2-antibody-11a9-ab252812'>ab252812</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

NCI-H1299 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

A549 (Human lung carcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution

Predicted band size: 97 kDa

Observed band size: 97 kDa

false

Exposure time: 100s

• IP

Supplier Data

Immunoprecipitation - Goat Anti-Rat IgG H&L (HRP) (AB205720)

non-muscle Myosin IIB/MYH10 was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney epithelial cell) whole cell lysate, 10µg with ab300647 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300647 at 1/1000 dilution. Goat Anti-Rat IgG (H+L), HRP) (ab205720) was used at 1/5000 dilution. Lane 1 : HEK-293T whole cell lysate, 10µg Lane 2 : ab300647 IP in HEK-293T whole cell lysate Lane 3 : Rat monoclonal IgG2a (ab18450) instead of ab300647 in HEK-293T whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-non-muscle Myosin IIB/MYH10 antibody [EPR22564-23] - Rat IgG2a (Chimeric) (<a href='/en-us/products/primary-antibodies/non-muscle-myosin-iib-myh10-antibody-epr22564-23-rat-igg2a-chimeric-ab300647'>ab300647</a>) at 1/30 dilution

All lanes:

HEK-293T (human embryonic kidney epithelial cell)

Secondary

All lanes:

Immunoprecipitation - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Observed band size: 250 kDa

false

Exposure time: 1s

• IP

Lab

Immunoprecipitation - Goat Anti-Rat IgG H&L (HRP) (AB205720)

VE Cadherin was immunoprecipitated from bEnd.3 whole cell lysate with ab91064 at 1/50 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab91064 at 1/1000 dilution.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-VE Cadherin antibody [BV13] - Intercellular Junction Marker (<a href='/en-us/products/primary-antibodies/ve-cadherin-antibody-bv13-intercellular-junction-marker-ab91064'>ab91064</a>) at 1/50 dilution

Lane 1:

bEnd.3 whole cell lysate at 10 µg

Lane 2:

bEnd.3 whole cell lysate

Lane 3:

Rat IgG1- Isotype Control instead of <a href='/en-us/products/primary-antibodies/ve-cadherin-antibody-bv13-intercellular-junction-marker-ab91064'>ab91064</a> in bEnd.3 whole cell lysate

Secondary

All lanes:

Immunoprecipitation - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/1000 dilution

Predicted band size: 90 kDa,120 kDa

false

Exposure time: 3s

• WB

Lab

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking/Dilution buffer : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256557).

All lanes:

Western blot - Anti-IDH1 antibody [RcMab-1] (<a href='/en-us/products/primary-antibodies/idh1-antibody-rcmab-1-ab256557'>ab256557</a>) at 1.479 µg/mL

Lane 1:

Wild type HAP1 whole cell lysate at 20 µg

Lane 2:

IDH1 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 5:

Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution

Predicted band size: 46 kDa

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking/Dilution buffer : 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 22715410).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab241544).

All lanes:

Western blot - Anti-SA1 antibody [SUSI63B] (<a href='/en-us/products/primary-antibodies/sa1-antibody-susi63b-ab241544'>ab241544</a>) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte) whole cell lysate at 10 µg

Lane 2:

MOLT-4 (human lymphoblastic leukemia T lymphoblast), whole cell lysate at 10 µg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 4:

A431 (human epidermoid carcinoma epithelial cell), whole cell lysate at 10 µg

Lane 5:

TF-1 (human Erythroleukemia erythroblast), whole cell lysate at 10 µg

Lane 6:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 10 µg

Lane 7:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution

Predicted band size: 144 kDa

false

Exposure time: 3s

• WB

Lab

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Exposure time : Lanes 1/3 : 3 seconds; Lane 2 : 15 seconds; Lane 4 : 1 second.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252878).

All lanes:

Western blot - Anti-WDR12 antibody [1B8] (<a href='/en-us/products/primary-antibodies/wdr12-antibody-1b8-ab252878'>ab252878</a>) at 1.235 µg/mL

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 2:

HEK-293 (human embryonic kidney epithelial cell), whole cell lysate at 10 µg

Lane 3:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 10 µg

Lane 4:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Predicted band size: 48 kDa

Observed band size: 47 kDa

false

• WB

Lab

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602).

Exposure time : Lanes 1-4 : 6 seconds, Lane 5：3 seconds, Lane 6：1 second.

Negative control : Raji

All lanes:

Western blot - Anti-CD3 antibody [CD3-12] (<a href='/en-us/products/primary-antibodies/cd3-antibody-cd3-12-ab11089'>ab11089</a>) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

Lane 2:

MOLT-4 (human lymphoblastic leukemia T lymphoblast), whole cell lysate at 20 µg

Lane 3:

Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate at 20 µg

Lane 4:

EL4 (mouse lymphoma T lymphocyte), whole cell lysate at 20 µg

Lane 5:

Mouse thymus tissue lysate at 20 µg

Lane 6:

Rat thymus tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Observed band size: 23 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : 15 secs.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252829).

All lanes:

Western blot - Anti-EWSR1/EWS antibody [21B1] (<a href='/en-us/products/primary-antibodies/ewsr1-ews-antibody-21b1-ab252829'>ab252829</a>) at 0.361 µg/mL

Lane 1:

K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

Ramos (human Burkitt's lymphoma B lymphocyte), whole cell lysate at 20 µg

Lane 4:

F9 (mouse embryonal carcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Predicted band size: 68 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

This data was developed using ab90806, the same antibody clone in a different buffer formulation.

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lane 1, 2 : 15 seconds; Lane 3 : 3 seconds; Lane 4 : 180 seconds.

All lanes:

Western blot - Anti-Pentraxin 3/PTX3 antibody [MNB1] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/pentraxin-3-ptx3-antibody-mnb1-bsa-and-azide-free-ab264487'>ab264487</a>) at 1/1000 dilution

Lane 1:

MDA-MB-231 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

Mouse placenta tissue lysate at 20 µg

Lane 3:

Rat placenta tissue lysate at 20 µg

Lane 4:

3T3-L1 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Predicted band size: 41 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

The expression profile weight observed is consistent with what has been described in the literature (PMID : 31634899).Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : 3 seconds

All lanes:

Western blot - Anti-Ferroptosis suppressor protein 1 antibody [6D8] (<a href='/en-us/products/primary-antibodies/amid-antibody-6d8-ab302673'>ab302673</a>) at 1/1000 dilution

Lane 1:

A375 (human malignant melanoma epithelial cell), whole cell lysate 20 µg

Lane 2:

HepG2 (human hepatocellar carcinoma epithelial cell), whole cell lysate 20 µg

Lane 3:

A549 (human lu carcinoma epithelial cell), whole cell lysate 20 µg

Lane 4:

T47D (human ductal breast epithelial tumor epithelial cell), whole cell lysate 20 µg

Lane 5:

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

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Observed band size: 40 kDa

false

Exposure time: 3s

• WB

Lab

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab6161 overnight at 4°C. Antibody binding was detected using ab205720, and visualised using ECL development solution ab133406.

true

Exposure time: 15s

• WB

Lab

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Negative control : HT-29, HepG2, A20.

The identity of the bands between 15 kDa and 37 kDa are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-CACHD1 antibody [15H10] (<a href='/en-us/products/primary-antibodies/cachd1-antibody-15h10-ab325744'>ab325744</a>) at 1/1000 dilution

Lane 1:

PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate at 40 µg

Lane 2:

SK-MEL-28 (human malignant melanoma cell ) whole cell lysate at 40 µg

Lane 3:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 4:

HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 5:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 40 µg

Lane 6:

F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 40 µg

Lane 7:

A20 (mouse reticulum sarcoma B lymphocyte) whole cell lysate at 40 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution

Observed band size: 170 kDa,36 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Low expression : SW480 and colo 205 (PMID : 31000598). Samples are non-boiled as boiling may cause protein aggregation. The expression profile observed is consistent with what has been described in the literature ( PMID : 32188872). Exposure time : 26 seconds

All lanes:

Western blot - Anti-xCT antibody [3A12] (<a href='/en-us/products/primary-antibodies/xct-antibody-3a12-ab300667'>ab300667</a>) at 1/1000 dilution

Lane 1:

A549 (human lung carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

SW480 (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 5:

COLO 205 (human colon; derived from metastatic site: ascites epithelial), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution

Observed band size: 55 kDa,35 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : 15 secs.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252820).

All lanes:

Western blot - Anti-BRD7 antibody [2D3] (<a href='/en-us/products/primary-antibodies/brd7-antibody-2d3-ab252820'>ab252820</a>) at 0.567 µg/mL

Lane 1:

C6 (rat glial tumor glial cell), whole cell lysate at 10 µg

Lane 2:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 10 µg

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 10 µg

Lane 4:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Predicted band size: 74 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab243153).

The molecular weight observed is consistent with what has been described in the literature (PMID : 3265124).

Exposure times : Lanes 1-3 : 3 seconds; Lane 4 : 3 minutes; Lane 5 : 15 seconds; Lane 6 : 44 seconds.

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Oct-2 antibody [9A2] (<a href='/en-us/products/primary-antibodies/oct-2-antibody-9a2-ab243153'>ab243153</a>) at 1/1000 dilution

Lane 1:

Ramos (human Burkitt's lymphoma B lymphocyte), whole cell lysate at 20 µg

Lane 2:

Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate at 20 µg

Lane 3:

Daudi (human Burkitt's lymphoma lymphoblast), whole cell lysate at 20 µg

Lane 4:

A20 (mouse reticulum sarcoma B lymphocyte), whole cell lysate at 20 µg

Lane 5:

Human tonsil tissue lysate at 20 µg

Lane 6:

Mouse spleen tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution

Predicted band size: 51 kDa

false

• WB

Lab

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Negative control : Raji (PMID : 11313386; PMID : 11054674).

Exposure time : lane 1, 2 : 10 seconds lane 3, 4 : 37 seconds

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252866).

All lanes:

Western blot - Anti-SH2D1A/SAP antibody [1D12] (<a href='/en-us/products/primary-antibodies/sh2d1a-sap-antibody-1d12-ab252866'>ab252866</a>) at 1/1000 dilution

Lane 1:

MOLT-4 (human lymphoblastic leukemia T lymphoblast), whole cell lysate at 20 µg

Lane 2:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

Lane 3:

Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate at 20 µg

Lane 4:

HDLM-2 (human Hodgkin lymphoma), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Predicted band size: 14 kDa

Observed band size: 14 kDa,40 kDa

false

• WB

Lab

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The identity of the higher MW band at approximately 37 kDa (in lane 1, 2) is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-CACHD1 antibody [15H10] (<a href='/en-us/products/primary-antibodies/cachd1-antibody-15h10-ab325744'>ab325744</a>) at 1/1000 dilution

Lane 1:

PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 50 µg

Lane 2:

PANC-1 transfected with siRNA specifically targeting CACHD1 whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution

Observed band size: 170 kDa,36 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

This data was produced using the same clone but in a different buffer, ab243140.

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Bax antibody [21C10] (<a href='/en-us/products/primary-antibodies/bax-antibody-21c10-ab243140'>ab243140</a>) at 1/5000 dilution

Lane 1:

HuT-78 (human Sezary syndrome cutaneous T lymphocyte), whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate at 20 µg

Lane 4:

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

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Predicted band size: 21 kDa

false

Exposure time: 1s

• WB

Lab

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking/Dilution buffer : 5% NFDM/TBST.

This image was produced using the same antibody clone but in a different formulation ab252825, PBS, sodium azide, glycerol and BSA.

All lanes:

Western blot - Anti-Dnmt1 antibody [2C1] (<a href='/en-us/products/primary-antibodies/dnmt1-antibody-2c1-ab252825'>ab252825</a>) at 0.353 µg/mL

Lane 1:

NCI-H1299 (human lung carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

HEK-293 (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

Lane 5:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/100000 dilution

Predicted band size: 183 kDa

false

Exposure time: 1s

• WB

Lab

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Samples are non-boiled as boiling may cause protein aggregates.

Exposure times : Lane 1 : 15 secs; Lane 2 : 92 secs.

Blocking/Dilution buffer : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252814).

All lanes:

Western blot - Anti-Amyloid Precursor Protein antibody [14D6] (<a href='/en-us/products/primary-antibodies/amyloid-precursor-protein-antibody-14d6-ab252814'>ab252814</a>) at 2.308 µg/mL

Lane 1:

Human hippocampus tissue lysate at 40 µg

Lane 2:

Rat hippocampus tissue lysate at 40 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution

Predicted band size: 86 kDa

Observed band size: 95 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lanes 1-2 : 15 secs; Lane 3 : 3 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252820).

All lanes:

Western blot - Anti-BRD7 antibody [2D3] (<a href='/en-us/products/primary-antibodies/brd7-antibody-2d3-ab252820'>ab252820</a>) at 0.567 µg/mL

Lane 1:

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

Lane 2:

PC-3 (human prostate adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Predicted band size: 74 kDa

false

• WB

Lab

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lane 1 : 3 minutes; Lanes 2-3 : 3 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256557).

All lanes:

Western blot - Anti-IDH1 antibody [RcMab-1] (<a href='/en-us/products/primary-antibodies/idh1-antibody-rcmab-1-ab256557'>ab256557</a>) at 1.479 µg/mL

Lane 1:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 10 µg

Lane 2:

Rat heart tissue lysate at 10 µg

Lane 3:

Human heart tissue lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution

Predicted band size: 46 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243148).

Blocking/diluting buffer and concentration : 5% NFDM/TBST.

Exposure time :

Lane 1 : 3 minutes
Lanes 2-5 : 3 seconds
Lane 6 : 5 seconds

Negative control : THP-1 and HL-60 (PMID : 10949928).

The molecular weight observed is consistent with what has been described in the literature (PMID : 9755173).

All lanes:

Western blot - Anti-BACH2 antibody [7A4] (<a href='/en-us/products/primary-antibodies/bach2-antibody-7a4-ab243148'>ab243148</a>) at 1/2000 dilution

Lane 1:

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

Lane 2:

Ramos (human Burkitt's lymphoma B lymphocyte), whole cell lysate at 20 µg

Lane 3:

THP-1 (human monocytic leukemia monocyte), whole cell lysate at 20 µg

Lane 4:

HL-60 (human Acute Promyelocytic Leukemia promyeloblast), whole cell lysate at 20 µg

Lane 5:

NAMALWA (human Burkitt's lymphoma B lymphocyte), whole cell lysate at 20 µg

Lane 6:

WEHI-231 (mouse B cell lymphoma B lymphocyte), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Predicted band size: 93 kDa

Observed band size: 130 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : 10 secs.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243136).

All lanes:

Western blot - Anti-MCL1 antibody [19C4-15] (<a href='/en-us/products/primary-antibodies/mcl1-antibody-19c4-15-ab243136'>ab243136</a>) at 1/1000 dilution

Lane 1:

SK-MEL-2 (human skin malignant melanoma) whole cell lysate at 20 µg

Lane 2:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 3:

C2C12 (mouse myoblasts myoblast) whole cell lysate at 20 µg

Lane 4:

EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/100000 dilution

Predicted band size: 37 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. This blot was developed using a high sensitivity ECL substrate. In Western blot, Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671) staining at 1/1000 dilution. Anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.Anti-His antibody (ab213204) staining at 1/5000 dilution. Exposure time : 3 minutes.

All lanes:

Western blot - Anti-ATG16L1 (phospho S278) antibody [EPR19016] - Rat IgG1 (Chimeric) (<a href='/en-us/products/primary-antibodies/atg16l1-phospho-s278-antibody-epr19016-rat-igg1-chimeric-ab309495'>ab309495</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector (vector control) containing a myc-His-tag whole cell lysate (untreated membrane) at 20 µg

Lane 2:

293T cells transfected with a human ATG16L1 expression vector containing a myc-His-tag whole cell lysate (untreated membrane) at 20 µg

Lane 3:

293T cells transfected with a human ATG16L1 (mutation S278A) expression vector containing a myc-His-tag whole cell lysate (untreated membrane) at 20 µg

Lane 4:

293T (human embryonic kidney epithelial cell) transfected with an empty vector (vector control) containing a myc-His-tag whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg

Lane 5:

293T cells transfected with a human ATG16L1 expression vector containing a myc-His-tag whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg

Lane 6:

293T cells transfected with a human ATG16L1 (mutation S278A) expression vector containing a myc-His-tag whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Observed band size: 68 kDa

false

• WB

Lab

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 seconds.

This image was produced using the same antibody clone but in a different formulation ab252869, PBS, sodium azide, glycerol and BSA.

All lanes:

Western blot - Anti-TPN antibody [7F6] (<a href='/en-us/products/primary-antibodies/tpn-antibody-7f6-ab252869'>ab252869</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

HeLa treated with 100 U/ml IFN gamma for 24 hours, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Predicted band size: 48 kDa

Observed band size: 47 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. This blot was developed using a high sensitivity ECL substrate. In Western blot, Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671) staining at 1/1000 dilution. anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 3 minutes.

All lanes:

Western blot - Anti-ATG16L1 (phospho S278) antibody [EPR19016] - Rat IgG1 (Chimeric) (<a href='/en-us/products/primary-antibodies/atg16l1-phospho-s278-antibody-epr19016-rat-igg1-chimeric-ab309495'>ab309495</a>) at 1/1000 dilution

Lane 1:

HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate (untreated membrane) at 20 µg

Lane 2:

HCT116 amino acid and serum starved in EBSS(HBSS) for 2h whole cell lysate (untreated membrane) at 20 µg

Lane 3:

HCT116 whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg

Lane 4:

HCT116 amino acid and serum starved in EBSS(HBSS) for 2h whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/1000 dilution

Observed band size: 68 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

The molecular weight observed was in consistency with the literature (PMID : 9299539).

Blocking buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Myelin Basic Protein antibody [12] - Oligodendrocyte Marker (<a href='/en-us/products/primary-antibodies/myelin-basic-protein-antibody-12-oligodendrocyte-marker-ab7349'>ab7349</a>) at 1/5000 dilution

All lanes:

Human cerebellum tissue lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution

Predicted band size: 33 kDa

Observed band size: 18.5 kDa

true

Exposure time: 1s

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

This data was developed using ab289888, the same antibody clone in a different buffer formulation.

Negative control : Jurkat (PMID : 18437383, 25168906

5% NFDM/TBST was used as a blocking and diluting buffer.

All lanes:

Western blot - Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Chimeric) (<a href='/en-us/products/primary-antibodies/granzyme-b-antibody-epr8260-rat-igg2a-chimeric-ab289888'>ab289888</a>) at 1/1000 dilution

Lane 1:

KARPAS-299 (human anaplastic large cell lymphoma), whole cell lysate at 20 µg

Lane 2:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

Lane 3:

SR (human pleural effusion lymphoblast), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Predicted band size: 28 kDa

Observed band size: 28 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking and diluting buffer : 5% NFDM/TBST

Negative control : MCF7 (HPA database)

Exposure time : 37 seconds

All lanes:

Western blot - Anti-Iba1 antibody [EPR16589] - Microglia marker - Rat IgG2a (Chimeric) (<a href='/en-us/products/primary-antibodies/iba1-antibody-epr16589-microglia-marker-rat-igg2a-chimeric-ab283346'>ab283346</a>) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte), 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-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Predicted band size: 16 kDa

Observed band size: 16 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

This data was developed using ab283347 the same antibody clone in a different buffer formulation.

Blocking and diluting buffer : 5% NFDM/TBST

Exposure time : 10 seconds

Negative control : Jurkat. (PMID : 27631416).

All lanes:

Western blot - Anti-PD-L2 antibody [CAL28] - Rat IgG2a (Chimeric) (<a href='/en-us/products/primary-antibodies/pd-l2-antibody-cal28-rat-igg2a-chimeric-ab283347'>ab283347</a>) at 1/1000 dilution

Lane 1:

HDLM-2 (human Hodgkin lymphoma) whole cell lysate at 20 µg

Lane 2:

Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Predicted band size: 31 kDa

Observed band size: 45-60 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

All lanes:

Western blot - Anti-SI-CLP antibody [1B8] (<a href='/en-us/products/primary-antibodies/si-clp-antibody-1b8-ab305176'>ab305176</a>) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte) transfected with scrambled siRNA control, whole cell lysate at 20 µg

Lane 2:

THP-1 transfected with siRNA specifically targeti SI-CLP, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution

Observed band size: 44 kDa

false

Exposure time: 62s

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking/Dilution buffer : 5% NFDM/TBST.

This image was produced using the same antibody clone but in a different formulation ab24736, PBS, sodium azide, glycerol and BSA.

All lanes:

Western blot - Anti-PPP2R1A antibody [6G3] (<a href='/en-us/products/primary-antibodies/ppp2r1a-antibody-6g3-ab24736'>ab24736</a>) at 0.785 µg/mL

Lane 1:

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

Lane 2:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

Lane 3:

Ramos (Human Burkitt’s lymphoma B lymphocyte), whole cell lysate at 20 µg

Lane 4:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 5:

C6 (rat glial tumor glial cell), whole cell lysate at 20 µg

Lane 6:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution

Predicted band size: 65 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lanes 1-3 : 5 secs; Lanes 4/5/7 : 3 secs; Lane 6 : 15 secs.

All lanes:

Western blot - Anti-MLKL antibody [3H1] (<a href='/en-us/products/primary-antibodies/mlkl-antibody-3h1-ab243142'>ab243142</a>) at 1/2000 dilution

Lane 1:

HUVEC (human umbilical vein endothelial cell), whole cell lysate at 10 µg

Lane 2:

HT-29 (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 4:

C6 (rat glial tumor glial cell), whole cell lysate at 10 µg

Lane 5:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 10 µg

Lane 6:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 10 µg

Lane 7:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/2000 dilution

Predicted band size: 54 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-RPA32/RPA2 antibody [4E4] (<a href='/en-us/products/primary-antibodies/rpa32-rpa2-antibody-4e4-ab252861'>ab252861</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

HUVEC (human umbilical vein endothelial cell ), whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

C6 (rat glial tumor glial cell), whole cell lysate at 20 µg

Lane 5:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Predicted band size: 29 kDa

false

• WB

Lab

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

ab181602 was used as a GAPDH loading control at a 1/200000 dilution.

In Western blot, anti-CBX1 antibody ab10811 was shown to bind specifically to CBX1. Target of interest was observed at 21kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in CBX1 knockout cell line (lane 2).

All lanes:

Western blot - Anti-CBX1 / HP1 beta antibody [MAC353] (<a href='/en-us/products/primary-antibodies/cbx1-hp1-beta-antibody-mac353-ab10811'>ab10811</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 2:

CBX1 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

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

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution

Observed band size: 21 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

The expression profile observed is consistent with what has been described in the literature (PMID : 19079332)

Blocking/diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-GBP1 antibody [1B1] (<a href='/en-us/products/primary-antibodies/gbp1-antibody-1b1-ab252832'>ab252832</a>) at 1/1000 dilution

Lane 1:

Untreated HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 2:

HeLa treated with 100U/ml IFN gamma (<a href='/en-us/products/proteins-peptides/recombinant-human-interferon-gamma-protein-active-ab9659'>ab9659</a>) for 24 hours, whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Predicted band size: 68 kDa

Observed band size: 67 kDa

false

Exposure time: 3s

• WB

Lab

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-CBX1 / HP1 beta antibody [MAC353] (<a href='/en-us/products/primary-antibodies/cbx1-hp1-beta-antibody-mac353-ab10811'>ab10811</a>) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution

Observed band size: 21 kDa

false

Exposure time: 125s

• WB

Lab

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lane 1, 2 : 15 seconds; Lane 3 : 3 seconds; Lane 4 : 180 seconds.

All lanes:

Western blot - Anti-Pentraxin 3/PTX3 antibody [MNB1] (<a href='/en-us/products/primary-antibodies/pentraxin-3-ptx3-antibody-mnb1-ab90806'>ab90806</a>) at 1/1000 dilution

Lane 1:

MDA-MB-231 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

Mouse placenta tissue lysate at 20 µg

Lane 3:

Rat placenta tissue lysate at 20 µg

Lane 4:

3T3-L1 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

Predicted band size: 41 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : 5 secs.

The negative control and the molecular weight of HCN1 observed is consistent with what has been described in the literature (PMID : 12786975).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252835).

All lanes:

Western blot - Anti-HCN1 antibody [7C3] (<a href='/en-us/products/primary-antibodies/hcn1-antibody-7c3-ab252835'>ab252835</a>) at 0.326 µg/mL

Lane 1:

Human brain tissue lysate at 20 µg

Lane 2:

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

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution

Predicted band size: 99 kDa

false

Exposure time: 5s

• WB

Lab

Western blot - Goat Anti-Rat IgG H&L (HRP) (AB205720)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Negative control : K562 (PMID : 27217538).

Exposure time : 59 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252813).

All lanes:

Western blot - Anti-AICDA antibody [5G9] (<a href='/en-us/products/primary-antibodies/aicda-antibody-5g9-ab252813'>ab252813</a>) at 1.264 µg/mL

Lane 1:

NAMALWA (human Burkitts lymphoma B lymphocyte), whole cell lysate at 10 µg

Lane 2:

Raji (human Burkitts lymphoma B lymphocyte), whole cell lysate at 10 µg

Lane 3:

K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 10 µg

Lane 4:

WEHI-231 (mouse B cell lymphoma B lymphocyte), whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/2000 dilution

Predicted band size: 23 kDa

Observed band size: 23 kDa

false