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
- GTX25658 - Provider product page
- Provider
- GeneTex
- Proper citation
- GeneTex Cat#GTX25658, RRID:AB_380629
- Product name
- PKC delta (phospho Ser664) antibody
- Antibody type
- Polyclonal
- Reactivity
- Human, Rat
- Host
- Rabbit
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Supportive validation
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- GeneTex (provider)
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- Experimental details
- Lysates prepared from K562 cells stimulated with PMA were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-12), or treated with lambda phosphatase (13), blocked with a 5% BSA-TBST buffer for one hour at room temperature, and incubated with PKCdelta [pS664] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 13), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphoserine-containing peptide (3), the phosphopeptide immunogen (4), or the phosphopeptides corresponding to other PKC isoforms (5-12). 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 PKCdelta [pS664] blocks the antibody signal. The peptide corresponding to PKCalpha [pS657] partially blocks the antibody signal, while those for PKCbetaII [pS660], and rat PKCdelta [pS662] block the antibody signal more thoroughly, indicating their respective reactivity with this antibody. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific