Key Specifications Table
|Key Applications||Detection Methods|
|Description||QCM Chemotaxis Cell Migration Assay, 24-well (5 µm), fluorimetric|
|Overview||Also available: Cell Comb™ Scratch Assay! Get biochemical data from a scratch assay! Click Here
The CHEMICON® QCM™ Chemotaxis 5 μm 24-well Migration Assay is performed in a Migration Chamber, based on the Boyden chamber principle. The 5 μm pore size of this assay's Boyden chambers is appropriate for studying monocyte/macrophage migration. The quantitative nature of this assay is useful for screening of pharmacological agents. Each kit provides sufficient materials for the evaluation of 24 samples.
Cell migration is a fundamental function of normal cellular processes, including embryonic development, angiogenesis, wound healing, immune response, and inflammationIn immune responses, monocyte migration is critical for providing immunological defense and eliminating infectious agents. Monocytes (macrophage precursors) circulate the blood until encountering specific chemotactic factors that initiate the migration process. Once engaged, monocytic cells transverse the vascular endothelium in a multi-step mechanism by: A) E-selectin mediated monocyte rolling B) cell adhesion C) transmigration into the subendothelium. During this migration and entering the tissue, monocytes develop/differentiate into mature macrophages, increasing in size, phagocytic activity, and proteolytic enzyme secretion. Once at the site of infection, mature macrophage respond to infection by phagocytosis, cytokine secretion, respiratory burst, T-cell antigen presentation, and digestive enzyme secretion.
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™ Chemotaxis 5 μm 24-well Migration Assay does not require cell labeling, scraping, washing, or counting. The 24-well inserts and homogenous fluorescence detection format allow for convenient screening and quantitative comparison of multiple samples.
In the Chemicon QCM™ Chemotaxis 5 μm 24-well Migration Assay, migratory cells on the bottom of the insert membrane are dissociated from the membrane when incubated with the Cell Detachment Solution. These cells are subsequently lysed and detected by the patented CyQUANT® GR dye (Molecular Probes). This green-fluorescent dye exhibits strong fluorescence enhancement when bound to cellular nucleic acids.
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™ Chemotaxis 5 μm 24-well Migration Assay utilizes a 5 μm pore size, which is appropriate for monocyte/macrophage migration. The system may be adapted to study different types of cell migration, including haptotaxis, random migration, chemokinesis, and chemotaxis.
The Chemicon QCM™ Chemotaxis 5μm 24-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.
|Application||The QCM Chemotaxis 5 um 24-well Migration Assay is performed in a Migration Chamber, based on the Boyden chamber principle.|
|Safety Information according to GHS|
|Storage and Shipping Information|
|Storage Conditions||Store kit components at 2° to 8°C, up to the expiration date provided on the kit.|
|Material Size||1 kit|
|Material Package||Contains 2 plates sufficient for 24 assays|
QCM Chemotaxis Cell Migration Assay, 24-well (5 µm), fluorimetric SDS
|Reference overview||Pub Med ID|
|Distinct signaling pathways for MCP-1-dependent integrin activation and chemotaxis|
Ashida, N, et al
J Biol Chem, 276:16555-16560 (2001) 2001
|Transmembrane motility assay of transiently transfected cells by fluorescent cell counting and luciferase measurement.|
Gildea, J J, et al.
BioTechniques, 29: 81-6 (2000) 2000
Current in vitro assays used in assessing tumor motility could be improved by the development of a simple technique that would facilitate studies of the impact of specific genes on pharmacologically altered chemotaxis. We developed a technique that improves on the classic transwell assay by using fluorescence and luminescence to assess chemotaxis. In this transient transfection system, co-transfection of a reporter construct and a gene with an unknown impact on motility are coupled with biochemical assays to quantitate the number of cells that have received a transferred gene, which subsequently crosses the membrane. This assay was found to be less variable than the conventional transwell chamber and is easily adaptable to studies of cell motility or cell invasion. We also demonstrate that this assay can detect the effect of both genetic and pharmacological inhibition of motility alone and in combination. It therefore has the potential to reveal additive or synergistic effects.
|Wound healing--aiming for perfect skin regeneration.|
Science, 276: 75-81 (1997) 1997
The healing of an adult skin wound is a complex process requiring the collaborative efforts of many different tissues and cell lineages. The behavior of each of the contributing cell types during the phases of proliferation, migration, matrix synthesis, and contraction, as well as the growth factor and matrix signals present at a wound site, are now roughly understood. Details of how these signals control wound cell activities are beginning to emerge, and studies of healing in embryos have begun to show how the normal adult repair process might be readjusted to make it less like patching up and more like regeneration.
|Transient functional expression of alphaVbeta 3 on vascular cells during wound repair.|
Clark, R A, et al.
Am. J. Pathol., 148: 1407-21 (1996) 1996
During early granulation tissue formation of wound repair, new capillaries invade the fibrin clot, a process that undoubtedly requires an interaction of vascular cells with the wound provisional matrix composed mainly of fibrin, fibronectin, and vitronectin. Integrin alphaVbeta3 is the vascular cell receptor for these wound-associated adhesive proteins. Therefore, we investigated the expression of this receptor on new capillaries of healing full-thickness cutaneous porcine wounds. During granulation tissue formation, alphaVbeta3 was expressed specifically on capillary sprouts invading the central fibrin clot whereas the closely related integrin alphaVbeta5 failed to localize to these cells. Cyclic peptides or antibody antagonists of alphaVbeta3 specifically inhibited granulation tissue formation in a transient manner during the period of invasive angiogenesis. Immunolocalization studies revealed that alphaVbeta3 became aggregated and lost from sprouting vessels after treatment with a peptide antagonist. In contrast, beta 1 integrins were not modulated by this treatment. Once granulation tissue filled the wound and invasive angiogenesis terminated, the alphaVbeta3 showed little or no expression in the granulation tissue microvasculature. These data demonstrate that integrin alphaVbeta3 plays a fundamental, but transient, role during invasive angiogenesis and granulation tissue formation in a healing wound.
|Simultaneous detection of free radical release and membrane current during phagocytosis|
Holevinsky, K O and Nelson, D J
J Biol Chem, 270:8328-8336 (1995) 1995