Using Western blot, a significant stimulation of NF-kappaB p65 phosphorylation at Ser536 is detected in HeLa cells treated with TNFalpha for 10 minutes (+) compared with untreated cells (-), while no change in total NF-kappaB p65 levels is observed.
Using the NF-kappaB p65 assay kits, a significant stimulation of NF-kappaB p65 phosphorylation at Ser536 is detected in HeLa cells treated with TNFalpha for 10 minutes (+) compared with untreated cells (-), while no change in total NF-kappaB p65 levels is observed.
Using the PhosphoTracer phospho-NF-kappaB p65 assay, the amount of lysate required to detect NF-kappaB p65 in TNFalpha-treated HeLa cell lysates was examined. Up to 50 µg of lysate/well was loaded into replicate wells of a PhosphoTracer microplate, and analysed for phospho-NF-kappaB p65. Phospho-NF-kappaB p65 was readily detected in less than 10 µg of lysate/well.
PhosphoTracer NF-kappaB p65 (pS536) ELISA Kit (ab119636) used in Sandwich ELISA.