Merck Millipore QCM Chemotaxis Cell Migration Assay 96-well Fluorimetric

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Introduction
Cell migration is a fundamental function of normal cellular processes, including embryonic development, angiogenesis, wound healing, immune response, and inflammation (1-2). One such process, leukocyte extravasation, is crucial for appropriate and effective immune response (3). Neutrophils normally exist in a resting state as they circulate though the body. However, upon interaction with small molecules known as chemoattractants, they rapidly respond with endothelial adhesion followed by emigration from the vasculature and chemotaxis to the site of inflammation (4-5). These chemoattractant receptors activate heterotrimeric GTP-binding proteins (G proteins) that initiate numerous elaborate signal transduction cascades, culminating in neutrophil migration and activation. Once at the site of inflammation, neutrophils respond with phagocytosis, superoxide generation, and the release of degradative enzymes.
Microporous membrane inserts are widely used for cell migration and invasion assays. The most widely accepted of which is the Boyden Chamber assay. However, current methods of analysis are time-consuming and tedious, involving cotton swabbing of non-migrated cells on the topside of insert, manual staining and counting. Recently a fluorescence blocking membrane insert was introduced to address these issues; however, this approach requires labeling of the cells with Calcein-AM and extensive washing to remove free Calcein before cell migration. The effect of this treatment on cell behavior/migration remains questionable.
The Chemicon QCM 96-well 3 μM Migration Assay does not require cell labeling, scraping, washing or counting. The 96-well insert and homogenous fluorescence detection format allows for large-scale screening and quantitative comparison of multiple samples (6).
In the Chemicon QCM 3 μM 96-well Migration Assay, migratory cells on the bottom of the insert membrane are dissociated from the membrane when incubated with Cell Detachment Buffer. These cells are subsequently lysed and detected by the patented CyQuant GR dye (Molecular Probes) (7). This green-fluorescent dye exhibits strong fluorescence enhancement when bound to cellular nucleic acids (8).
Most migration assays utilize an 8 μm pore size, as this is appropriate for most cell types, e.g. epithelial and fibroblast cells. The Chemicon QCM 3 μM 96-well Migration Assay utilizes a 3 μm pore size, which is appropriate for leukocyte migration. The system may be adapted to study different types of cell migration, including haptotaxis, random migration, chemokinesis, and chemotaxis.
The Chemicon QCM 3 μM 96-well Migration Assay provides a quick and efficient system for quantitative determination of various factors on cell migration, including screening of pharmacological agents, evaluation of integrins or other adhesion receptors responsible for cell migration, or analysis of gene function in transfected cells.
In addition, Chemicon also provides QCM 8μM 96-well Chemotaxis Cell Migration (ECM510) and Invasion Assays (ECM555) (8 μm pore), 24-well insert Cell Migration and Invasion Assay Systems, CytoMatrix Cell Adhesion strips coated with ECM proteins, or anti-integrin antibodies, and QuantiMatrix ECM protein ELISA kits.
Application:
The Chemicon QCM 96-well 3μM Migration Assay is ideal for studying chemotaxis cell migration. The quantitative nature of this assay is especially useful for large scale screening of pharmacological agents. The 3 μm pore size of this assay's Boyden chambers is appropriate for studying leukocyte migration. Each kit provides sufficient materials for the evaluation of 96 samples.
The Chemicon QCM 3μM 96-well Migration Assay is intended for research use only; not for diagnostic applications.