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
- GTX25686 - Provider product page
- Provider
- GeneTex
- Proper citation
- GeneTex Cat#GTX25686, RRID:AB_380652
- Product name
- ERK5 (phospho Thr218/Tyr220) antibody
- Antibody type
- Polyclonal
- Reactivity
- Human, Mouse, Simian
- Host
- Rabbit
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Supportive validation
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- GeneTex (provider)
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- Experimental details
- Extracts of HEK293 cells transiently transfected with plasmids expressing ERK5 kinase domain (ERK5kin) and constitutively activated MEK5D-D were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer overnight at 4 ¢J, then incubated with the ERK5 (phospho Thr218) antibody (GTX25686) for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1), the non-phosphopeptide corresponding to the phosphopeptide immunogen (2), a generic phosphothreonine-containing peptide (3), a generic phosphotyrosine-containing peptide (4), the phosphopeptide derived from the corresponding region of ERK1&2 (5), or the phosphopeptide immunogen (6). The data show that while there is some cross-reactivity with ERK1&2, only the phosphopeptide corresponding to ERK5 [pTpY218/220] completely blocks the antibody signal, demonstrating the specificity of the antibody. NOTE: The antibody signal appears at ~50 kDa as this is the molecular weight of the transiently transfected ERK5 kinase domain.