One-step analysis of protein complexes in microliters of cell lysate
- 21 October 2005
- journal article
- Published by Springer Nature in Nature Methods
- Vol. 2 (11), 833-835
- https://doi.org/10.1038/nmeth802
Abstract
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.Keywords
This publication has 12 references indexed in Scilit:
- Chemical Inhibitors when Timing Is Critical: A Pharmacological Concept for the Maturation of T Cell ContactsChemBioChem, 2005
- IL-2 and IL-15 receptor α-subunits are coexpressed in a supramolecular receptor cluster in lipid rafts of T cellsProceedings of the National Academy of Sciences, 2004
- Minimal Requirement of Tyrosine Residues of Linker for Activation of T Cells in TCR Signaling and Thymocyte DevelopmentThe Journal of Immunology, 2003
- Mechanism of Recruitment of WASP to the Immunological Synapse and of Its Activation Following TCR LigationMolecular Cell, 2002
- Identification of the Minimal Tyrosine Residues Required for Linker for Activation of T Cell FunctionJournal of Biological Chemistry, 2001
- Mapping the Zap-70 phosphorylation sites on LAT (linker for activation of T cells) required for recruitment and activation of signalling proteins in T cellsBiochemical Journal, 2001
- Mapping the Zap-70 phosphorylation sites on LAT (linker for activation of T cells) required for recruitment and activation of signalling proteins in T cellsBiochemical Journal, 2001
- A Novel and Robust Homogeneous Fluorescence-Based Assay Using Nanoparticles for Pharmaceutical Screening and DiagnosticsSLAS Discovery, 2000
- GRB2 and phospholipase C-gamma 1 associate with a 36- to 38-kilodalton phosphotyrosine protein after T-cell receptor stimulation.Molecular and Cellular Biology, 1994
- Fluorescence correlation spectroscopy with high count rate and low background: analysis of translational diffusionEuropean Biophysics Journal, 1993