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Acad acetate acrylamide activity aliquots amount antigen antisera aqueous assay bacteria bacteriophage T4 binding Biol cDNA cell lines chloramphenicol Cl pH cloned gene codon column containing culture denatured dithiothreitol DNA polymerase EDTA EDTA pH 8.0 efficiency elution encoding Escherichia coli ethanol ethidium bromide eukaryotic extract film foreign protein fragment hybridization immunoglobulin immunoprecipitation Incubate ligation lysate 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 nucleotides P.O. Box pellet phosphate plasmid plate PMSF polyclonal precipitate prepared Proc promoter protein of interest radioactive radiolabeled reaction reagent remove restriction enzyme Resuspend room temperature sample SDS gel-loading buffer SDS-polyacrylamide gel sequences serum sodium sodium azide staining sterile stock solution stored supernatant target antigen target protein Telephone termini transcription transfection transfer transient expression volume wash western blotting