44-716G

antibody from Invitrogen Antibodies

Targeting: ASAP2 CENTB3, DDEF2, KIAA0400, PAP, SHAG1

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

44-716G

Provider

Invitrogen Antibodies

Product name

Phospho-ASAP2 (Tyr724) Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Peptide Competition - Mutant Analysis Lysates prepared from COS cells overexpressing human wild-type PAG3 alone (1) cotransfected with human wild-type PAG3 and mouse Pyk2 (2-5), or mutant (Y724F/Y763F) PAG3 and mouse Pyk2 (6) were resolved by SDS-PAGE on an 8% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer for one hour at room temperature, then incubated with PAG3 (pY724) antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 2, 6), the non-phosphopeptide corresponding to the immunogen (3), a generic phosphotyrosine-containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F (ab’)2 anti-rabbit IgG HRP conjugate (Product # ALI4404) in 3% BSA-TBST buffer, and bands were detected using the Pierce SuperSignal™ method. The data show that only the peptide corresponding to PAG3 (pY724) blocks the signal, and the signal is absent in the mutant extracts, demonstrating the specificity of the antibody. Cell extracts were kindly provided by Dr. Hisataka Sabe, Osaka Bioscience Institute, Osaka, Japan.