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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
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- Product number
- GTX18031 - Provider product page
- Provider
- GeneTex
- Proper citation
- GeneTex Cat#GTX18031, RRID:AB_423035
- Product name
- PAG3 (phospho Tyr724) antibody
- Antibody type
- Polyclonal
- Reactivity
- Human
- Host
- Rabbit
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Supportive validation
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- GeneTex (provider)
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- Experimental details
- 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 in 3% BSA-TBST buffer, and bands were detected. 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.
