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Download Hybridoma Technology in the Biosciences and Medicine by Zelig Eshhar (auth.), Timothy A. Springer (eds.) PDF

By Zelig Eshhar (auth.), Timothy A. Springer (eds.)

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Try to avoid growth of many clones per well by using low seeding density. 3. Do not allow dense growth! Dilute and transfer the hybridomas to fresh wells when required. 4. Aminopterin can be omitted 7-10 days after fusion. Phase out HAT with several changes of HT medium over a period of at least 10 days. 5. Freeze positive hybridomas as soon as possible. 6. Clone positive hybridomas as soon as possible. 7. Always repeat subcloning of positive clones. Monoclonal Antibody Strategy and Techniques 4.

Culture supernatants have been assayed for antibody activity in a variety of tests, such as standard or solid phase radioimmunoassay (RIA) and enzymelinked immunosorbent assay (ELISA), that will be described in detail in the following sections. Other methods that are less frequently used are immunoflourescence, immunoprecipitation, neutralization of biological activity mediated by the antigen, complement-mediated lysis of target cells or coated red cells, hemagglutination, and others. These are described in detail elsewhere.

87-89 Similarly, for MAbs intended to serve for affinity purification of antigens from complex mixtures, it is helpful to select for a MAb that binds the antigenic molecule and can dissociate from it under conditions that will retain the activity of both the antigen and antibody. We have designed and used such screening procedures for the derivation of MAbs to human interferons. 12 - 14 Other factors that determine the design of the screening procedure are related to the purity, accessibility, and physical form of the antigen.

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