Metabolic labeling & Immunoprecipitation

A metabolic labeling and IP protocol by Jun Takagi from Springer lab at CBR Institute for Biomedical Research.

Cells incorporate 35S-methionine or cysteine during the protein synthesis. Thus it is essential to use Met,Cys-free medium and dialyzed FCS during the labeling. Short period of incubation with 35S-methionine or cysteine will result in radiolabeling (pulse), and additional incubation with excess concentration of unlabeled Met+Cys (chase) is needed for complex glycoproteins like integrins to get expressed as a maturated form.

Last update 09-May-2005, Rating n/a of 0 votes.