Anti-cIAP2 antibody [E40] (ab32059) at 1/50000 dilution (Purified) + Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysates at 15 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 72 kDa
Observed band size: 72 kDa

Anti-cIAP2 antibody [E40] (ab32059) at 1/1000 dilution (unpurified) + Daudi cell lysate.

Predicted band size: 72 kDa
Observed band size: 72 kDa

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast carcinoma tissue sections labeling cIAP2 with purified ab32059 at 1/600 dilution (1.12 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

Flow Cytometry analysis of Raji (Human Burkitt's lymphoma B lymphocyte) cells labeling cIAP2 with purified ab32059 at 1/70 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

ab32059 (purified) at 1/30 dilution (2ug) immunoprecipitating cIAP2 in Daudi whole cell lysates.
Lane 1: Daudi (Human Burkitt's lymphoma lymphoblast) whole cell lysates 10ug
Lane 2 (+): ab32059 & Daudi whole cell lysates
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32059 in Daudi whole cell lysates
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.

ab32059 (unpurified) at a 1/50 dilution staining cIAP2 in human normal spleen, using Immunohistochemistry, Paraffin Embedded Tissue.

cIAP2 was immunoprecipitated using 0.5mg Daudi whole cell extract, 5µg of Rabbit polyclonal to cIAP2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Daudi whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70^oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab32059 (unpurified).
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 72kDa; cIAP2