Anti-cIAP1 antibody [EPR4673] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-cIAP1 antibody [EPR4673] - BSA and Azide free (AB226031)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human spleen tissue sections labeling cIAP1 with Purified ab108361 at 1 : 500 dilution (2.82 �g/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108361).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-cIAP1 antibody [EPR4673] - BSA and Azide free (AB226031)

This IHC data was generated using the same anti-cIAP1 antibody clone, EPR4673, in a different buffer formulation (cat# ab108361).

ab108361, at 1/50, staining cIAP1 in paraffin-embedded Human spleen tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-cIAP1 antibody [EPR4673] - BSA and Azide free (AB226031)

This WB data was generated using the same anti-cIAP1 antibody clone, EPR4673, in a different buffer formulation (cat# ab108361).

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : cIAP1 knockout HAP1 cell lysate (20 μg)
Lane 3 : HeLa cell lysate (20 μg)
Lane 4 : Jurkat cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab108361 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab108361 was shown to specifically react with cIAP1 when cIAP1 knockout samples were used. Wild-type and cIAP1 knockout samples were subjected to SDS-PAGE. ab108361 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10,000 dilution respectively and incubated overnight at 4C. Blots were developed withGoat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-cIAP1 antibody [EPR4673] (<a href='/en-us/products/primary-antibodies/ciap1-antibody-epr4673-ab108361'>ab108361</a>)

Predicted band size: 70 kDa

false

• WB

Lab

Western blot - Anti-cIAP1 antibody [EPR4673] - BSA and Azide free (AB226031)

This data was developed using the same antibody clone in a different buffer formulation (ab108361).

Lanes 1- 2 : Merged signal (red and green). Green - ab108361 observed at 70 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab108361 was shown to react with cIAP1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265896 (knockout cell lysate ab257372) was used. Wild-type HeLa and BIRC2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab108361 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-cIAP1 antibody [EPR4673] (<a href='/en-us/products/primary-antibodies/ciap1-antibody-epr4673-ab108361'>ab108361</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

BIRC2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human BIRC2 (cIAP1) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-birc2-ciap1-knockout-hela-cell-line-ab265896'>ab265896</a>)

Predicted band size: 70 kDa

Observed band size: 70 kDa

false