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By David A. Cheresh

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Fibronectin is adsorbed in PBS-ABC for 30 min at room temperature, the excess removed, and the plate washed 1Â with PBS-ABC. The coverslips are blocked for 30 min with 1% BSA (heat inactivated at 56 for 30 min), washed 3Â with PBS-ABC or HEPES adhesion buffer and stored in adhesion buffer up to 3 hours. Other extracellular matrix proteins will have different adsorption profiles but are roughly in the same range as fibronectin (approximately threefold). , 2006). Plastic is rougher than glass and adsorbs about twice the level of protein (Fig.

2000; Reyes and Garcia, 2003). To detach a spread fibroblast would require $300,000 g. Hence, this approach is not that practical as a cell detachment assay and the inability of this method to detach cells that are attached by more than $10 bonds means that the data look very like the plate-and-wash assays. Therefore, in order to measure cell detachment forces, a method that applied more force was required. , 1997). The advantage of hydrodynamic shear is that it can apply much higher forces, and the forces are to a large extent distributed over the cell surface rather than applied at a point.

P. MATHER, AND GORDON H. SATO VOLUME 199. Cumulative Subject Index Volumes 168–174, 176–194 VOLUME 200. Protein Phosphorylation (Part A: Protein Kinases: Assays, Purification, Antibodies, Functional Analysis, Cloning, and Expression) Edited by TONY HUNTER AND BARTHOLOMEW M. SEFTON VOLUME 201. Protein Phosphorylation (Part B: Analysis of Protein Phosphorylation, Protein Kinase Inhibitors, and Protein Phosphatases) Edited by TONY HUNTER AND BARTHOLOMEW M. SEFTON VOLUME 202. Molecular Design and Modeling: Concepts and Applications (Part A: Proteins, Peptides, and Enzymes) Edited by JOHN J.

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