Antibodies, A Laboratory Manual, Second Edition, Edited by Edward A. Greenfield



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Collecting Tissue Culture Supernatants

(Protocol summary only for purposes of this preview site)

When collecting tissue culture supernatants for antibodies, allow the individual cultures to grow until the hybridomas die. This will allow the collection of higher-titer supernatants. In general, antibodies are resistant to the proteases that are released from dying cells, so allowing the cells to die should not affect the quality of the antibodies. If extraneous IgG molecules will alter any of the assays for which the supernatants are being prepared, use medium with fetal bovine serum or use serum-free medium. The yield of this method is 2050 g of antibody/mL of supernatant.

Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition

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