A wide range of fixed and live cell imaging protocols, including FRAP and FRET techniques, are detailed here:

Trinkle-Mulcahy, L., Chusainow, J., Lam, Y.W., Swift, S. and Lamond, A.I. Visualization of intracellular PP1 targeting through transiently and stably expressed fluorescent protein fusions. Methods in Molecular Biology. Vol.. 365:133-54; Protein Phosphatase Protocols, Ed. G. Moorhead, Humana Press, 2006.

Details about the the SILAC-based affinity purification/quantitative proteomics approach can be found in our recent methods paper:

Trinkle-Mulcahy L., Boulon S., Lam Y.W., Urcia R., Boisvert F.M., Vandermoere F., Morrice N.A., Swift S., Rothbauer U., Leonhardt H. and Lamond A.I. Identifying specific protein interaction partners using quantitative mass spectrometry and bead proteomes. J Cell Biol. 183:223-39, 2008.

..and also in these PDF files:

For details on sourcing reagents and preparing the media, download our SILAC Reagent Protocol.

For details on covalently coupling antibodies to Protein G sepharose, download our Covalent Coupling Protocol.

For details on cell fractionation and preparation of whole cell vs. nuclear and cytoplasmic lysates, download our Cell Fractionation Protocol.

For details on trypsin digestion of gel slices for MS analysis, download our In Gel Digestion Protocol.




 











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