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Innocyte™ Cell Migration Assay  | | | - a convenient assay for screening prospective antimigratory compounds,
or studying the effect of various chemotactic substances - | | | | | | Intended Use:
The Innocyte™ Cell Migration Assay has been designed to study the effects of various drugs or agents on< cell motility and to identify chemoattractant molecules (chemotactic migration). The 8 mm pore-size of the membrane ensures that non-specific, random migration is minimized. Cell migration through the membrane is quantified by staining the cells that attach to the lower side of the membrane with a fluorescent dye. The staining and dislodgement of the cells from the lower side of the membrane is performed simultaneously. Notes: The membrane is not coated with an extracellular matrix protein, and therefore not suitable for the study of haptotactic migration. The assay is not intended for leukocyte migration experiments, as the study of leukocytes requires membranes with a pore-size smaller than 8 mm. | | | Background Information:
Cell migration is crucial for embryonic development, the inflammatory immune response, wound repair, and tumor formation, and metastasis.1 Recent findings highlight the importance of tension, the actin and myosin filament network, the Rho/Rac family of signaling molecules, and microtubules in cell migration.2, 3 The coordination of these complex processes is the challenge facing cells that are on the move. The regulation of focal adhesion and focal complex turnover is critical for the continued remodeling and reorganization of adhesion contacts during cell migration, and migratory defects have been reported in cells lacking Src family kinases, focal adhesion kinase (FAK), and calpain.4-6 When the cells in a tumor develop the ability to migrate out of the initial tumor and invade their surroundings, they become very dangerous. Some cells may migrate into the circulatory system and move to new locations (metastasis), where they then form a new (secondary) tumor. This change in the cells comprising a tumor, from sedentary to migratory, represents a serious problem, one which may be solved with a better understanding of the migratory process itself.
| | | Principle of the Assay:
The Innocyte™ Cell Migration Assay is provided in a convenient 96-well format suitable for screening prospective antimigratory compounds or studying the effect of various chemotactic substances. Cell migration is quantified by detaching and labeling the migrated cells, which cling to the bottom of the membrane, with a fluorescent dye (1 step). There is no need for manual cell scraping or cell counting. | | |
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| InnoCyte™ Cell Migration Assay, 96-well |
96-well plate |
3-5 h |
Adherent cells |
CBA010 |
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| | | Kit Contents:
96-well Cell Migration Chamber: 1 sterile multi-well plate with cell culture inserts and tray
Additional Tray (needed in cell detachment step): 1 plastic tray
Cell Detachment Buffer: 20 ml in a plastic bottle
Calcein-AM Solution: 70 ml in an amber vial
Latrunculin A Solution (Antimigratory Compound): 25 ml in a clear vial (1 mM) | | | Additional Information:
Figure 1: Human umbilical venous endothelial cells (HUVECs) were allowed to migrate across a membrane insert toward media in the presence or absence of chemotactic components for 4 hours at 37°C in a 6% CO2 incubator (chemotactic migration). Approximately 50,000 cells were loaded per well in triplicate. Migrated cells were subsequently dislodged and fluorescently labeled from the underside of the membrane. Latrunculin A, a powerful inhibitor of actin polymerization, was added to the cell suspension immediately before loading the cells on the migration plate. 
Figure 2: A highly migratory capacity is exhibited by human fibrosarcoma cells (HT-1080). HT-1080 cells were allowed to migrate across a membrane insert toward media in the presence or absence of serum for 3 hours at 37°C in a 6% CO2 incubator (chemotactic migration). Approximately 60,000 cells were loaded per well, in triplicate. Migrated cells were subsequently dislodged and fluorescently labeled from the underside of the membrane. 
| | |  We also offer other Assay Kits for the study of Angiogenesis - View Angiogenesis Kit Selection Guide | | |
Check out the excellent technical articles and large product offering in our Angiogenesis and Tumor Metastasis Brochure!
View PDF I Request your free copy | |
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