GTX25796

antibody from GeneTex

Targeting: PRKCG MGC57564, PKCC, PKCG, PKCγ, SCA14

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Flow cytometry (Data presented on provider website)

Antibody Data

Product number

GTX25796

Provider

GeneTex

Proper citation

GeneTex Cat#GTX25796, RRID:AB_380691

Product name

PKC gamma (phospho Thr655) antibody

Antibody type

Polyclonal

Reactivity

Human, Mouse

Host

Rabbit

Western blot

Supportive validation

Submitted by

GeneTex (provider)

Main image

Experimental details

Lysates prepared from HeLa cells stimulated with PMA were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-11) or treated with lambda phosphatase (12), blocked with a 3% low-fat milk-TBST buffer overnight at 4¢XC, and incubated with 0.50 ?g/ml PKCgamma [pT655] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 11, 12), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphothreonine containing peptide (3), the phosphopeptide immunogen (4), or, the phosphopeptide corresponding to the immunogen from other PKC isoforms (5-10). After washing, membranes were incubated with goat F(ab)2 anti-rabbit IgG alkaline phosphatase and signals were detected. The data show that the peptide corresponding to PKCgamma [pT655] blocks the antibody signal while the peptides corresponding to PKC isoforms betaI [pT642], delta [pS645], epsilon [pT710] and eta [pT655] do not completely block the antibody signal, thereby demonstrating the specificity of the antibody. However, peptides corresponding to PKC isoforms alpha [pT638] and betaII [pT641] do block the antibody signal, indicating that the PKCgamma [pT655] antibody may cross-react with their respective sites. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.