Fol. Biol. 2009, 55, 153-158
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.
Keywords
integrins, tyrosine kinases, cell adhesion, phosphotyrosine, cytoskeleton.
Funding
This work was supported by grants of the Ministry of Education, Youth and Sports of the Czech Republic (Research Center grant LC06061, project MSM0021620858).
References
Copyright
This is an open-access article distributed under the terms of the Creative Commons Attribution License.
