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
- GTX25798 - Provider product page
- Provider
- GeneTex
- Proper citation
- GeneTex Cat#GTX25798, RRID:AB_380693
- Product name
- PKC eta (phospho Thr655) 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 stimulated with PMA were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-9) or treated with lambda phosphatase (10), blocked with a 5% BSA-TBST buffer overnight at 4¢XC, and incubated with 0.35 ?g/ml PKCeta [pT655] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 10), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphothreonine containing peptide (3), the phosphopeptide immunogen (4), or, the phosphopeptide corresponding to the immunogen from other PKC isoforms (5-9). After washing, membranes were incubated with goat F(ab)2 anti-rabbit IgG HRP-conjugate and bands were detected. The data show that the peptide corresponding to PKCeta [pT655] blocks the antibody signal. The antibody signal was not blocked by the peptides corresponding to PKC isoforms alpha [pT638], betaI [pT642], betaI [pT641], gamma [pT655] and zeta [pT560], thereby demonstrating the specificity of the antibody. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.