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



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Metabolic Labeling of Antigens with [35S]Methionine

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[35S]Methionine or a combination of [35S]methionine and [35S]cysteine are the most commonly used radioactive precursors used for labeling proteins (Chandra et al. 1998; Jung and Cashman 1999; Bonifacino 2001; Biermann et al. 2002; Girao and Pereira 2003). Although a variety of amino acids containing either [3H] or [14C] are commercially available, [35S] is easier to detect than either [3H] or [14C]. The intracellular presence of methionine is generally less than that of many other amino acids. Therefore, exogenous [35S]methionine added to the medium is readily taken up by the cell and incorporated into newly translated proteins. The addition of a mixture comprising [35S]methionine and [35S]cysteine to the medium is particularly useful for increasing the specific labeling of proteins, particularly if the target protein has relatively few methionines.

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

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