GTX25798

antibody from GeneTex

Targeting: PRKCH PKC-L, PKCL, PRKCL

Western blot (Supportive data in Antibodypedia)

Flow cytometry (Data presented on provider website)

Antibody Data

Product number

GTX25798

Provider

GeneTex

Proper citation

GeneTex Cat#GTX25798, RRID:AB_380693

Product name

PKC eta (phospho Thr655) antibody

Antibody type

Polyclonal

Reactivity

Human

Host

Rabbit

Western blot

Supportive validation

Submitted by

GeneTex (provider)

Main image

Experimental details

Lysates prepared from Jurkat cells stimulated with PMA were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-9) or treated with lambda phosphatase (10), blocked with a 5% BSA-TBST buffer overnight at 4¢XC, and incubated with 0.35 ?g/ml PKCeta [pT655] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 10), 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 (5-9). After washing, membranes were incubated with goat F(ab)2 anti-rabbit IgG HRP-conjugate and bands were detected. The data show that the peptide corresponding to PKCeta [pT655] blocks the antibody signal. The antibody signal was not blocked by the peptides corresponding to PKC isoforms alpha [pT638], betaI [pT642], betaI [pT641], gamma [pT655] and zeta [pT560], thereby demonstrating the specificity of the antibody. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.