These 96-well microplate-based assays are designed to accelerate the screening process for compounds that influence chemotaxis. The Cell Migration Assay measures the number of cells traversing a porous membrane, while the Cell Invasion Assays monitor cell movement through extracellular matrices. Invasive migration is a fundamental function underlying cellular processes such as angiogenesis, embryonic development, immune response, metastasis, and invasion of cancer cells. The Cell Invasion Assays offer a flexible, standardized, high-throughput format for quantitating the degree to which invasive cells penetrate a barrier consisting of basement membrane components in response to chemoattractants and/or inhibiting compounds.
These assay kits contain all the reagents necessary to investigate cell migration through Basement Membrane Extract (BME), Laminin I, Collagen I, or Collagen IV. Cultrex Cell Invasion Assay Kits include a simplified 96-well Boyden chamber with an 8 μm polyethylene terephthalate (PET) membrane that must be coated with the extracellular matrix by the end-user.
These ready-to-use assay kits contain a 96-well Boyden chamber with a PET membrane pre-coated with BME at either a low, medium or high density, to investigate cell invasion in vitro. The ideal coating density can be determined using an optimization assay that contains a 96-well Boyden chamber with wells pre-coated with three different BME concentrations.
Cell Invasion Assay. This assay employs a simplified Boyden chamber-like design that consists of two chambers separated by a filter coated with BME or different extracellular matrix components. The cell suspension is placed in the top chamber and incubated in the presence of test media containing specific chemoattractants in the bottom chamber. Cells migrate from the top chamber through the coated filter pores to the bottom of the filter. Detection of cell invasion is quantified using Calcein AM. Cell dissociation/Calcein AM solution is placed in the bottom chamber to dissociate the migrating cells from the filter. Calcein AM is internalized by the cells, and intracellular esterases cleave the acetomethylester (AM) moiety. Free Calcein fluoresces brightly and is used to quantitate the number of cells that have invaded or migrated by comparison with a standard curve.
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