Antibody data
- Antibody Data
- Antigen structure
- References [0]
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- Validations
- Western blot [1]
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Validation data
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- Product number
- GTX24774 - Provider product page
- Provider
- GeneTex
- Proper citation
- GeneTex Cat#GTX24774, RRID:AB_380419
- Product name
- eIF4E (phospho Ser209) antibody
- Antibody type
- Polyclonal
- Reactivity
- Human, Mouse, Rat, Rabbit
- Host
- Rabbit
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Supportive validation
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- GeneTex (provider)
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- Experimental details
- Extracts of HeLa cells were resolved by SDS-PAGE on a 4-20% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer for one hour at room temperature, either left untreated (1-4) or treated with lambda phosphatase (5), and then incubated with the EIF-4E [pS209] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1,5), the non-phosphopeptide corresponding to the phosphopeptide immunogen (2), a generic phosphoserine-containing peptide (3), or the phosphopeptide immunogen (4). After washing, the membrane was incubated with goat F(ab)2 anti-rabbit IgG alkaline phosphatase and signals were detected. The data show that only the phosphopeptide corresponding to EIF-4E [pS209] blocks the antibody signal, demonstrating the specificity of the antibody. The data also show that phosphatase stripping eliminates the signal, further verifying that the antibody is phospho-specific.