Cell Adherence Assay (LTI) General and nice Protocol for cell adherence assay. Proteins are coated on microtiter plates and cells are added; after the nonbound cells are washed away, the number of bound cells can be semi-quantitated by staining them and reading the absorbance in a microtiter plate reader.
Cell Adherence Inhibition Assay (LTI) General protocol--Monoclonal antibodies to individual integrin subunits have been characterized for their ability to interrupt normal adhesion to ECM ligands, and can be used to determine factors that may affect the cell activity. Alternatively, many extracellular proteins contain the peptide sequence RGD (Arg-Gly-Asp) in different configurations that confer specificity to different integrin receptors, and the addition of the soluble peptide can also interrupt the normal adhesion process. Either monoclonal antibody or RGD peptide is added along with the cells during the standard adhesion assay, and the inhibitory effect can thus be semi-quantitated.
Dynamic Flow Assay in a Parallel Plate Flow Chamber (Glycotek) Flow assays allow visualization of cell adhesion under well-defined wall shear stress. The visualization of the different events of cell adhesion can be quantified by selective image acquisition and subsequent image processing. Flow assays are uniquely suited to the investigation of adhesive events which occur very rapidly in a time scale shorter than that of most static adhesion assays. In addition, events subsequent to the initial events can be studied such as cell stabilization and spreading giving some insight into the kenetics of particular cell-cell or cell-substrate adhesive behavior.
Measurement of Cell Adhesion Under Static Conditions (Glycoteck) This protocol introduces the use of a liquid-filled wash chamber that separates unbound cells by gravity thereby eliminating uncontrolled shear forces and passage of adherent cells through a liquid/air interface. This protocol is also useful for assaying molecules that promote or inhibit cell adhesion.
Carbohydrate-Specific Adhesion of Intact Cells to Resolved Glycolipids on TLC Plates (Glycoteck) his procedure allows detection of specific cell adhesion to glycolipids resolved on TLC plates. The steps include: (1) HPTLC of glycoconjugates, (2) coating the chromatogram with polymer, (3) preblocking the chromatogram to reduce non-specific cell binding, (4) mounting the chromatogram in the acrylic chamber, (5) adding labeled cells, (6) incubating, (7) removal of non-adherent cells by centrifugation, and (8) fixation of adherent cells for detection.
"Cells on Gels" (The Cell Biology and Cytoskeleton Group, HMS) This protocol describes method for culture cell on a thin polyacrylamide-based,collagen-coated flexible substrate. By maintaining a constanttotal concentration of acrylamide while usr/localying the concentrationof bis-acrylamide, it's able to obtain a series of chemicallyidentical substrates with a wide range of flexibility. By usingimaging techniques, cells' response to differencesin substrate flexibility can be detected.
