Antibody data
- Antibody Data
- Antigen structure
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- Comments [0]
- Validations
- Western blot [1]
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Validation data
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- Product number
- GTX12862 - Provider product page
- Provider
- GeneTex
- Proper citation
- GeneTex Cat#GTX12862, RRID:AB_373050
- Product name
- Syk (phospho Tyr323) antibody
- Antibody type
- Polyclonal
- Reactivity
- Human, Mouse
- Host
- Rabbit
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Supportive validation
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- GeneTex (provider)
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- Experimental details
- Extracts of Jurkat cells untreated (1) or treated with 10 mM H2O2 for 3 minutes (2-6) were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was either left untreated (1-5) or treated with YOP phosphatase (6), then blocked with a 3% milk-TBST buffer for one hour at room temperature, and incubated with the Syk [pY323] antibody for two hours at room temperature in a 3% milk-TBST buffer, following prior incubation with: no peptide (1, 2, 6), the non-phosphopeptide corresponding to the phosphopeptide immunogen (3), a generic phosphotyrosine-containing peptide (4), or the phosphopeptide immunogen (5). After washing, the membrane was incubated with goat F(ab¡¦)2 anti-rabbit IgG HRP conjugate and signals were detected. The data show that only the phosphopeptide corresponding to Syk [pY323] blocks the antibody signal, demonstrating the specificity of the antibody. The data also show the up-regulation of phosphorylation upon treatment with H2O2 in this cell system and that phosphatase stripping eliminates the signal, further verifying that the antibody is phospho-specific.