Analyzing integrin-dependent adhesion.
This unit describes methods for the analysis of integrin-ligand binding in both cell-based assays and solid-phase assays. A major application of cell adhesion assays is in investigating whether a certain cell type can adhere to a specific adhesive substrate, and, if so, which receptors are involved. Particularly if the substrate is a matrix component (e.g., fibronectin), members of the integrin family are likely to play a dominant role in adhesion. Procedures are described here for assessing which integrins are involved in this process. A detailed analysis of ligand recognition by individual integrins can be performed using a solid-phase receptor-ligand binding assay. The unit also contains support protocols for integrin purification and coupling of antibodies to Sepharose for use in this purification, as well as for biotinylation of integrin ligands to be used in the solid-phase assay.
Mould
A Paul
AP
University of Manchester, Manchester, United Kingdom.
D000818:Animals; D000906:Antibodies / Q000032:analysis / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology*; D006801:Humans; D016023:Integrins / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification / Q000502:physiology*
D000906:Antibodies; D016023:Integrins