This is a knockout-validated antibody summary, based on the publication "Genetic Compensation Triggered by Mutant mRNA Degradation", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Figure 1. Western blot analysis of RELA and ACTB levels in Rela K.O. cells transfected with empty vectors (control) or plasmids encoding wt RELA. From [1].

Mouse monoclonal IgG2b

Company: Cell Signaling

Antibody: NF-κB p65

Catalog number: 6956

Summary: Mouse monoclonal IgG2b against a synthetic peptide corresponding to residues near the carboxy terminus of human NF-κB protein. Reacts with human, mouse, rat, hamster, monkey, mink, bovine, dog, and pig. Suitable for western blot, immunoprecipitation, immunohistochemistry (paraffin), immunofluorescence/immunocytochemistry, flow cytometry and chromatin IP.

Western blot

Rela KO mouse embryonic stem cells transfected with empty vectors (control) or plasmids encoding wt RELA. were lysed in modified RIPA buffer (150 mM NaCl, 50 mM TrisHCl pH 7.4, 1% IGEPAL, 0.1% sodium deoxycholate, 1 mM EDTA) supplemented with protease inhibitors (cOmplete ULTRA Mini, Roche) and PMSF.

5% BSA (sigma) for 1 hr.

1:1,000 dilution overnight at 4°C.

1:10,000 dilution anti-mouse IgG-HRP (Thermo, 31430) for 1 hr at RT.

ECL (Clarity Western ECL Substrate, Biorad) and imaged using a ChemiDoc MP system (Biorad).