Antibody Usage in the Lab
Laura Caponi, Paola Migliorini
Springer Berlin Heidelberg, Aug 15, 1999 - Science - 154 pages
A handy lab manual that permits quick and easy access to the techniques commonly used to analyse antibody specificity. Some of the most useful immunological techniques based on antibodies are described, including ELISA, immunoblotting and immunoprecipitation protocols that provide useful tools for recognising immunological specificities, and basic immunofluorescence and immunohistochemistry procedures for the in situ identification of antigens. The topics are discussed from a practical point of view, describing the theoretical basis of each technique and sample protocols plus a troubleshooting guide. Attention is focused on the various aspects of the protocols described thus providing the reader with the maximum possible information on each technique.
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900 ml distilled add distilled water adjust to pH alcohol amino acid anti antibody e.g. antigen assay avidin biotin biotinylated blocking blotting bovine serum albumin cells Centrifuge coating commercially available concentration containing detection dilution Dissolve distilled water EDTA ELISA enzyme epitope Ethanol final volume fixation fixative flow cytometer fluorochrome gelatine hour immunoblot immunoprecipitation Incubate labelled antibody LAURA CAPONI Magnetic stirrer membrane minutes ml distilled water ml double-distilled water molecules monoclonal antibodies Na2HPO4 NaCl in 800 NaN3 nitrocellulose Note paraffin pellet peptides Pisa plate polyclonal precipitate prepared primary antibody procedure proteins protocol reaction reagent room temperature sample saturating agent second antibody slides sodium spot staining buffer stock solution Store at 4°C streptavidin Subprotocol supernatant suspension techniques tion tissue transfer Tris-buffered saline TTBS tube Tween 20 University of Pisa volume of 1000 Wash three Wash twice Washing step