R&D Systems offers a range of aseptically prepared microplates pre-coated with extracellular matrix (ECM) proteins for culturing adherent cell lines and quantifying cell adhesion.
Many proteins of the ECM interact with cells via cell surface integrin family receptors. The resulting focal contacts are important for the maintenance of tissue architecture and for supporting a variety of cellular processes. ECM protein/integrin binding may initiate a complex network of signal transduction cascades that, depending on the context, plays an important role in cell spreading, migration, proliferation, and differentiation during embryogenesis, wound healing, and tumor development.
Kits are available containing 96-well microplates pre-coated with Fibronectin (Catalog # CWP001) or Vitronectin (Catalog # CWP003). These aseptic plates may be used to promote cell attachment during culture of adherent cell lines.
Microplates pre-coated with ECM proteins can also be used as a tool to quantify cell adhesion. Cell suspensions are simply applied to pre-coated microplates and centrifuged to promote interaction with the plate surface. Non-adherent cells are gently removed during PBS washes. The percentage of adherent cells is determined by measuring Calcein AM fluorescence before and after the wash steps.
*This assay requires additional reagents; Calcein AM (Catalog # 4892-010-K) and PBS with calcium and magnesium.
|Figure 1. R&D Systems Extracellular Matrix (ECM) Protein Coated Plate assay template (left). As indicated by text in wells, rows A and H are left uncoated to serve as controls, and rows B through G are coated with ECM protein. The entire plate is then blocked with 1% Bovine Serum Albumin Fraction V (BSA). In this assay, cells were either unstimulated (burgundy) or stimulated (orange) with 10 ng/mL PMA for 30 minutes and applied to R&D Systems Human Fibronectin Coated Plates (Catalog # CWP001) in the pattern shown. Cells were labeled with calcein-AM and adhesion to the plates was measured as a percentage of total cells applied (right). All bars depict average and standard error values obtained from three plates.|
Coat your own plates with R&D Systems recombinant ECM proteins.
An alternative approach to promote cell adhesion.