Smac antibody | knockout validation | Abcam ab32023

This is a knockout-validated antibody summary, based on the publication "Mitochondrial permeabilization engages NF-κB-dependent anti-tumour activity under caspase deficiency", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Antibody information

Rabbit monoclonal IgG

Company: Abcam

Antibody: Smac

Catalog number: ab32023

Summary: Rabbit monoclonal IgG against a synthetic peptide within Human Smac/ Diablo (C terminal). Reacts with human, mouse, and rat. Suitable for western blot, immunoprecipitation, immunohistochemistry (paraffin), immunofluorescence (immunocytochemistry), and flow cytometry.

Validation Method

Western blot

Sample

Wild-type or SMAC/OMI-/- E1A/ras transformed MEF cells. Cells were lysed in NP-40 lysis buffer (1% NP-40, 1mM EDTA, 150mM NaCl, 50 mM Tris pH 7.4, 1 mM PMSF and complete protease inhibitors (Roche)).

Primary incubation

1/1000 dilution overnight in 4°C.

Secondary incubation

HRP-linked secondary antibodies.

Detection
Figure

Wild-type or SMAC/OMI-/- E1A/ras transformed MEFs were treated with ABT-737 (10 μM) and UMI-77 (10 μM) and Q-VD-OPh (30 μM) for 4 h. Cell lysates were

immunoblotted for indicated proteins. Please see Figure 4f in the article [1].

Clone note

The same clone (Y12) is sold as Abcam ab32023, ab199317; EMD Millipore 04-578.

References
  1. Giampazolias E, Zunino B, Dhayade S, Bock F, Cloix C, Cao K, et al. Mitochondrial permeabilization engages NF-κB-dependent anti-tumour activity under caspase deficiency. Nat Cell Biol. 2017;19:1116-1129 pubmed publisher