Cell pellets were lysed in urea buffer, protein concentration was measured via micro BCA assay (Thermo), and peptides were prepared and immunoprecipitated as described25,57. Approximately 250 × 106 total CD4+ T cells at the start of stimulation yielded ∼3 mg of total peptide. Peptides ...
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Cell pellets were lysed in urea buffer, protein concentration was measured via micro BCA assay (Thermo), and peptides were prepared and immunoprecipitated as described25,57. Approximately 250 Â 106 total CD4 þ T cells at the start of stimulation yielded B3 mg of total peptide. Peptides ...
Cell lysates were size-fractioned in 10–12% SDS-polyacrylamide before transfer to Hybond TM-C Extra membranes (GE Healthcare, Buckinghamshire, UK). Membranes were blocked and incubated overnight at 4 ◦C with the antibodies. Anti-EGFR, anti-p 1068EGFR, anti-Bax, anti-Bcl2, anti-PARP,...