Key Spec Table
|Key Applications||Detection Methods|
|Description||QCM Chemotaxis Cell Migration Assay, 24-well (3µm), colorimetric|
|Overview||Also available: Cell Comb™ Scratch Assay! Get biochemical data from a scratch assay! Click Here
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. 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. 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. The Boyden Chamber system uses a hollow plastic chamber, sealed at one end with a porous membrane. This chamber is suspended over a larger well which may contain medium and/or chemoattractants. Cells are placed inside the Chamber and allowed to migrate through the pores, to the other side of the membrane. Migratory cells are then stained and counted.
Most migration assays utilize an 8 µm pore size, as this is appropriate for most cell types, e.g. epithelial and fibroblast cells. The Millipore 3 µm QCM™ Chemotaxis Assay 24-well- colorimetric utilizes a 3 µm pore size, which is appropriate for leukocyte migration. Cells migrated toward chemoattractants were measured by WST-1 (cell viability stain), and quantitated by spectrophotometers. The system may be adapted to study different types of cell migration, including haptotaxis, random migration, and chemokinesis.
The Millipore QCM™ 3 µm Chemotaxis Assay 24-well-colorimetric 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 continues to provide numerous migration, invasion, and adhesion products including:
• QCM™ 8µm 24-well Chemotaxis Cell Migration Assays (ECM508, 509)
• QCM™ 5µm 24-well Chemotaxis Cell Migration Assay (ECM507)
• QCM™ 8µm 96-well Chemotaxis Cell Migration Assay (ECM510)
• QCM™ 5µm 96-well Chemotaxis Cell Migration Assay (ECM512)
• QCM™ 3µm 96-well Chemotaxis Cell Migration Assay (ECM515)
• QCM™ 96-well ECMatrix™ Cell Invasion Assay (ECM555)
• QCM™ 96-well Collagen-based Cell Invasion Assay (ECM556)
|Materials Required but Not Delivered||1. 96-well ELISA plate.
2. Precision pipettes: sufficient for aliquoting cells.
3. Harvesting buffer: EDTA or trypsin based cell detachment buffer, or other cell detachment formulations as optimized by individual investigators. Millipore's ready-to-use non-mammalian detachment solution, Accutase™ (Cat. No. SCR005) can also be used.
Note: Trypsin cell detachment buffer maybe required for strongly adherent cell lines, but can strip cell surface proteins. Allow sufficient time for cell receptor recovery.
4. Tissue culture growth medium appropriate for subject cells, such as RPMI-1640 containing 10% FBS.
5. Chemoattractants (eg. 10% FBS) or pharmacological agents for addition to culture medium, if screening is desired.
6. Sterile PBS or HBSS to wash cells.
7. Distilled water.
8. Low speed centrifuge and tubes for cell harvesting.
9. CO2 incubator appropriate for subject cells.
10. Hemocytometer or other means of counting cells.
11. Trypan blue or equivalent viability stain.
12. Spectrophotometer (microplate reader).
13. Sterile cell culture hood.
|Safety Information according to GHS|
|Storage and Shipping Information|
|Storage Conditions||The two Sterile 3 µm 24-well Cell migration plate assembly and forceps are stored at 2°-8°C. WST-1 reagent and Electro Coupling Solution are stored at -20°C.|
|Material Size||1 plate|
|Material Package||24 wells|