Rabbit polyclonal to Histone H1 phospho T18 (1/1000) against recombinant Histone H1, incubated with control lysate (lanes 1, 3, 5, 7, 9, 11, 13) or insect cell lysate containing active cyclin E / CDK2 complexes (lanes 2, 4, 6, 8, 10, 12, 14).Mutant Histone H1 proteins were made by site-directed mutagenesis of 5 phosphorylation cyclin/cdk phosphorylation concensus sites (T18, T146, T154, S172, S187). Lanes 3-12 contain mutant histone H1 with only one wild-type cyclin/cdk phosphorylation concensus site and lanes 13-14 containst mutant Histone H1 with all 5 sites mutated to Alanine.Lanes 1-2    :  wt Histone H1Lanes 3-4    :  T146A, T154A, S172A, S187A (wt site at T18)Lanes 5-6    :  T18A, T154A, S172A, S187A (wt site at T146)Lanes 7-8    :  T18A, T146A, S172A, S187A (wt site at T154)Lanes 9-10  :  T18A, T146A, T154A, S187A (wt site at S172)Lanes 11-12:  T18A, T146A, T154A, S172A (wt site at S187)Lanes 13-14:  T18A, T146A, T154A, S172A, S187AAlejandro Contreras, Baylor College of Medicine