Cell Signaling Technology
Phospho-p38 MAPK (Thr180/Tyr182) Antibody
9211
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Many signals via MKK3, MKK6, activate p38 alpha MAPK by phosphorylation at Thr180 and Tyr182, which in turn phosphorylates a many other proteins.The levels of Thr180 and Tyr182 phosphorylated p38 alpha is used as a measure of activation of stress activated MAPK pathway.
Western Blot
HePG2 cells
Primary antibody (1:1000) in dilution buffer (TBST with 5% BSA) with gentle agitation overnight at 4°C.
TBST with 5% w/v nonfat dry milk
Incubated the membrane with the Goat Anti-Rabbit IgG, H & L Chain Specific Peroxidase Conjugate (Cat No: 401315) 1:10000 diluted in 10 ml of blocking buffer with gentle agitation for 1 hr at room temperature.
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Incubated the membrane with appropriate amount of Luminata Crescendo Western HRP substrate (WBLUR0100 Merckmillipore) for 5 min at room temperature. Drained the membrane of excess developing solution (without letting it to dry) wrapped in transparent plastic sheet and exposed the x-ray film for 1 min.
Hydrogen peroxide treatment to cells is a well known activator of p38 alpha and downstream pathway . The antibody detects the activated p38-alpha when treated with hydrogen peroxide to the cells.
This antibody is also used in the published article PMID: 26660341
Works well without detecting much background or unspecific proteins.
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Useful antibody for determination of p38 pathway activation.