One-step analysis of protein complexes in microliters of cell lysate.
We present 'mix and measure' procedures for the analysis of protein complexes in microliters of crude human and mouse cell lysates using fluorescence correlation and crosscorrelation spectroscopy. We labeled interacting endogenous proteins by indirect immunofluorescence with all primary and secondary reagents added in one step. Especially for the screening of compounds interfering with interactions that depend on signaling-induced posttranslational modifications, the approach represents a major advance over existing protocols.
Stoevesandt
Oda
O
Interfaculty Institute for Cell Biology, Department of Molecular Biology, University of Tübingen, Auf der Morgenstelle 15, 72076 Tübingen, Germany.
Köhler
Karsten
K
Fischer
Rainer
R
Johnston
Ian C D
IC
Brock
Roland
R
D000818:Animals; D002457:Cell Extracts / Q000737:chemistry*; D002460:Cell Line; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods*; D006801:Humans; D051379:Mice; D046912:Multiprotein Complexes / Q000032:analysis* / Q000737:chemistry*; D012680:Sensitivity and Specificity; D015398:Signal Transduction; D013057:Spectrum Analysis / Q000379:methods*
D002457:Cell Extracts; D046912:Multiprotein Complexes