Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
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Validation data
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- Product number
- 44-232G - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-Zap-70 (Tyr315, Tyr319) Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Storage
- -20°C
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Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Peptide Competition and Phosphatase Treatment. 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 (Product # ALI4404) and bands were detected using the Pierce SuperSignal™ method. 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.