Detection of human IRS-1, phosphorylated at Tyr896, by immunoblotting. Samples: Whole cell lysate from CHO-T cells transfected with human IRS-1, stimulated with 50 nM insulin (for 5 min) (all lanes) and left untreated (lanes 1-5, 7) or treated with CD45 (tyrosine phosphatase) (lanes 6, 8). Primary antibody: PhosphoDetect™ Anti-IRS-1 (pTyr896) Rabbit pAb (Cat. No. GF1003) pre-incubated without peptide (lane 1, 7, 8), with a generic phosphotyrosine-containing peptide (lane 2), non-phosphopeptide corresponding to the immunogen (lane 3), or phosphopeptide immunogen (lane 4) (1:1000) and pan IRS-1 antibody (lanes 5, 6). Detection: chemiluminescence.
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