Fibronectin-replating experiment: procedure and analysis.
In this review protocols are described for studying protein tyrosine kinase signalling upon integrin-mediated cell adhesion. We have outlined detailed procedures for fibronectin-replating experiment, biochemical examination of the phosphotyrosine content of cellular proteins by immunoblotting using phosphorylation-specific antibodies or immunoprecipitation and analysis with general phosphotyrosine antibodies. Despite great advances that were made toward optimizing the described procedures, all these methods still remain in many respects an art, given the plentiful of variables and the extent to which the optimum conditions vary from one experimental condition to the other. Examples of performed experiments using the described procedures thus also include notes regarding variability of approaches based on experimental conditions.
Rösel
D
D
Charles University in Prague, Faculty of Science, Department of Cell Biology, Prague, Czech Republic.
Půta
F
F
Brábek
J
J
D000818:Animals; D045163:Antibodies, Phospho-Specific / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D005353:Fibronectins / Q000378:metabolism*; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism
D045163:Antibodies, Phospho-Specific; D005353:Fibronectins; D051416:Focal Adhesion Protein-Tyrosine Kinases