Anti Mer2, phospho Ser30

Western blot using Rockland's affinity purified anti S.cerevisiae Mer2 pS30 antibody shows detection of phosphorylated Mer2 in whole cell extracts.？ Cells were either wild type (+) or contained mer2 deletions (D). Extracts were prepared from cells 4hr after initiation of meiosis.？ Proteins were obtained using TCA precipitation.？ The primary antibody was used at a 1:7,500 dilution.？ Secondary antibody was used at 1:5,000 dilution.？ Personal Communication. Michael Lichten, NIH, CCR, Bethesda, MD.

Western blot using Rockland's affinity purified anti-S.cerevisiae Mer2 pS30 antibody shows detection of phos-phorylated Mer2 but not phosphatase treated or mutant cells.？ Lane 1 contains Mer2-myc protein detected in wild type cells after first immunoprecipitating the protein using anti-myc antibody.？ Cells were harvested 4 h after the initiation of meiosis and therefore contain mostly phosphorylated Mer2.？ Lane 2 contains the same preparation after treatment with phosphatase.？ Lane 3 contains Mer2-S30A protein as a phosphorylation control.？ This antibody appears to be specific for phosphorylated Mer2 at the S30 position with negligible cross reactivity against unphosphor-ylated protein.？ The primary antibody was used at a 1:5,000 dilution.？ Personal Communication. Michael Lichten, NIH, CCR, Bethesda, MD.