GTX25785

antibody from GeneTex

Targeting: PRKCB PKCB, PKCβ, PRKCB1, PRKCB2

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Flow cytometry (Data presented on provider website)

Antibody Data

Product number

GTX25785

Provider

GeneTex

Proper citation

GeneTex Cat#GTX25785, RRID:AB_380685

Product name

PKC beta 2 (phospho Thr641) antibody

Antibody type

Polyclonal

Reactivity

Human, Rat

Host

Rabbit

Western blot

Supportive validation

Submitted by

GeneTex (provider)

Main image

Experimental details

Peptide Competition and Phosphatase Treatment: Lysates prepared from K562 cells stimulated with PMA were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either treated with lambda phosphatase (1) or, left untreated (2-5), blocked with a 5% BSA-TBST buffer for two hours at room temperature, and incubated with primary antibody for two hours at room temperature in a 3% BSATBST buffer, following prior incubation with: no peptide (1, 2), the nonphosphopeptide corresponding to the immunogen (3), a generic phosphothreoninecontaining peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab)2 anti-rabbit IgG HRP conjugated and bands were detected. The data show that the peptide corresponding to PKC beta2 [pS641] blocks the antibody signal. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.

Supportive validation

Submitted by

GeneTex (provider)

Main image

Experimental details

PMA-treated K652 cell lysates were incubated with anti-PKC-beta II [pT641] after prior incubation with: no peptide (1, 2), the non-phosphopeptide (3), a generic [pT] peptide (4), or, the phosphopeptide immunogen (5). Peptide competition data show antibody phospho-site specificity. Lane 1 was treated with l phosphatase prior to immunoblot. Absence of signal further demonstrates antibody specificity.

Validation comment

WB