Antibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
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Validation data
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- Product number
- GTX12869 - Provider product page
- Provider
- GeneTex
- Proper citation
- GeneTex Cat#GTX12869, RRID:AB_367105
- Product name
- ZAP70 (phospho Tyr315/Tyr319) 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 Jurkat cells left untreated (1) or treated with H2O2 (2-6) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-6) or treated with YOP phosphatase (7), blocked with a 5% BSA-TBST buffer for one hour at room temperature, and incubated with ZAP-70 [pYpY315/319] 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) the phosphopeptide immunogen (5) or, the phosphopeptide corresponding to ZAP-70 [pY315] (6). After washing, membranes were incubated with goat F(ab)2 anti-rabbit IgG HRP conjugate and bands were detected. The data show that only the peptide corresponding to ZAP-70 [pYpY315/319] completely blocks the antibody signal. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific