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acetate acrylamide activity Adjust the volume aliquots amino acids ammonium persulfate amount antigen antisera aqueous assay autoclaving bacteria bacteriophage T7 binding bovine serum albumin cDNA cell lines cloned gene coli column culture deionized H2O denatured described on pages dilution Dissolve dithiothreitol DNA polymerase EDTA EDTA pH 8.0 efficiency elution Escherichia coli ethanol eukaryotic expression extract foreign protein fragment glycerol hybridization immunoglobulin immunoprecipitation Incubate lysate lysis buffer mammalian cells medium method mg/ml microfuge tube minutes at 4°C minutes at room mixture monoclonal antibodies mRNA NaCl Natl nitrocellulose nitrocellulose filter nucleic acid pellet phosphate plasmid plate PMSF polyclonal precipitate prepared promoter protein of interest radioactive radiolabeled reaction reagent Remove restriction enzyme Resuspend room temperature sample SDS gel-loading buffer SDS-polyacrylamide gel sequence sodium sodium azide sterile stock solution strain supernatant target antigen target protein termini transcription transfection Transfer Tris Cl pH vectors wash western blotting