A wide range of fixed
and live cell imaging protocols, including FRAP and FRET techniques,
are detailed here:
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
For details on cell fractionation and preparation of whole cell
vs. nuclear and cytoplasmic lysates, download our Cell
For details on trypsin digestion of gel slices for MS analysis,
download our In Gel Digestion