GTX24767

antibody from GeneTex

Targeting: RAF1 c-Raf, CRAF, Raf-1

Western blot (Supportive data in Antibodypedia)

Immunoprecipitation (Data presented on provider website)

Flow cytometry (Data presented on provider website)

Antibody Data

Product number

GTX24767

Provider

GeneTex

Proper citation

GeneTex Cat#GTX24767, RRID:AB_380415

Product name

Raf1 (phospho Ser621) antibody

Antibody type

Polyclonal

Reactivity

Human, Rat

Host

Rabbit

Western blot

Supportive validation

Submitted by

GeneTex (provider)

Main image

Experimental details

Antibody-Peptide Competition and Mutant Analysis Immunoprecipitates of Hek293 cells overexpressing wild-type c-Raf and stimulated with 50 ng/mL EGF for 3 minutes (1-4) or c-Raf mutant S621A (5) were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer overnight at 4¢J, then incubated with the c-Raf [pS621] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: the phosphopeptide immunogen (1), a generic phosphoserine-containing peptide(2), the non-phosphopeptide corresponding to the phosphopeptide immunogen (3), or no peptide (4, 5). The inset shows the relative amount of total c-Raf protein in the wild-type vs. the S621A mutant extracts via c-Raf pan antibody. The data show that only the phosphopeptide corresponding to c-Raf [pS621] blocks the antibody signal and that the antibody does not recognize the S621A mutant, demonstrating the specificity of the antibody.