WB and IP of Rabbit anti-IL-7-Receptor-alpha-chain-pY449 antibody. Lane 1: thymocyte D1 cells. Lane 2: thymocyte D1 cells treated with IL7 (50 ng/ml). IP: with anti-Phosphotyrosine conjugated to Protein G agarose. Primary antibody: IL7pY449 antibody at 1:10000 for overnight at 4°C. Secondary antibody: HRP-conjugated goat anti-rabbit antibody at 1:10,000 for 45 min at RT and ECL detection. Block: 5% BLOTTO overnight at 4°C.