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Janus kinase (JAK)2 tyrosine kinase is constitutively associated with IL-5Rα, and JAK1 tyrosine kinase with IL-5Rβc

Chemicals

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Cell pellets bearing His 6 -GATE16 were resuspended in 50 ml of buffer NLB (10 mM K 2 HPO 4 , 300 mM NaCl, 20 mM KCl, 10 mM imidazole, 5 mM 2-mercaptoethanol, pH 8.0), lysed and clarified as above and then loaded onto a 5-ml Ni-NTA column (Bio-Rad), washed with two column volumes of buffer NLB, and eluted over 20 column volumes in a linear gradient of buffer NLB with imidazole increasing from 10 mM to 1 M