Current Protocols in Molecular Biology, Volume 2
Frederick M. Ausubel, Roger Brent, Robert E. Kingston, David D. Moore, J. G. Seidman, Kevin Struhl
John Wiley & Sons, Nov 14, 1988 - Science - 1600 pages
Current Protocols in Molecular Biology (CPMB), the first in the Current Protocols family of laboratory manuals, is the international standard by which all other laboratory manuals are judged. Originally published in 1987, CPMB now boasts of three looseleaf volumes (soon to expand to four volumes). Basic methods for DNA preparation and isolation, library screening, and sequencing have been joined by more advanced procedures detailing DNA-protein interactions, yeast manipulation, and phosphorylation analyses. From basics to the cutting edge, CPMB is the resource you need for successful experiments. Carefully edited, step-by-step protocols replete with material lists, expert commentaries, and safety and troubleshooting tips ensure that you can duplicate the experimental results in your own laboratory. Quarterly updates, which are filed into the looseleaf, keep the set current with the latest developments in molecular biology methods. The initial purchase includes one year of updates and then subscribers may renew their annual subscriptions. Current Protocols publishes a family of laboratory manuals for bioscientists, including Immunology, Human Genetics, Protein Science, Cytometry, Cell Biology, Neuroscience, Pharmacology, and Toxicology.
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INTO MAMMALIAN CELLS
7A Isotopic Assay for Reporter Gene Activity 9 7 1
7C Use of the A Victoria Green Fluorescent Protein to Study Protein
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acrylamide addition aliquots Alternate Protocol amino acid ammonium persulfate Analysis of Proteins antigen ascites assay Basic Protocol binding buffer capillary cell lines centrifuge chromatography cloning column complex concentration conjugate containing culture detection determined dilution efficiency electroporation elution enzyme epitope ethanol expression extract filter flow rate fluorescence fraction gel electrophoresis gene gradient HPLC hybridoma immunoprecipitation Incubate infection interactions ion-exchange ions isotyping luciferase Mammalian Cells mass spectrometry matrix medium membrane method mg/ml microcentrifuge microcentrifuge tube molecular molecules monoclonal antibodies nitrocellulose optimal peak pellet peptides and proteins phosphate pipet plasmid plate Prepare procedure produce purification reagents Reagents and Solutions recipe recombinant remove reporter gene residues resuspend retroviral room temperature sample SDS-PAGE selection separation Sepharose sequence serum sodium soluble solvent specific staining step Store substrate supernatant Support Protocol techniques tion tissue titer transfection transfer tube unit vector viral virus volume wash