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QCM ECMatrix Cell Invasion Assay, 24-well (8 μm), fluorimetric
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Cell Based Assays
Millipore offers a significant portfolio of well-published, quantitative and optimized live cell, whole-cell, and cell-based activity assays. Study Apoptosis, Angiogensis, Adhesion and more.
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Description:
QCM ECMatrix Cell Invasion Assay, 24-well (8 μm), fluorimetric
Trade Name:
QCM
Chemicon (Millipore)
Qty/Pk:
24 assays
Product Overview:
Also available: Cell Comb™ Scratch Assay! Get biochemical data from a scratch assay! Click Here)
Introduction
Invasion through the extracellular matrix (ECM) is an important step in tumor metastasis. Cancer cells initiate invasion by adhering to and spreading along the blood vessel wall. Proteolytic enzymes, such as MMP collagenases, dissolve tiny holes in the sheath-like covering (basement membrane) surrounding the blood vessels to allow cancer cells to invade (1).
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-invaded 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 invasion. The effect of this treatment on cell behavior/invasion remains questionable.
The Chemicon® QCM™ 24-well Invasion Assay does not require cell labeling, scraping, washing or counting. The 24-well insert and homogenous fluorescence detection format allows for large-scale screening and quantitative comparison of multiple samples.
In the Chemicon® QCM™ 24-well Invasion Assay, invaded 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) (2-3). This green-fluorescent dye exhibits strong fluorescence enhancement when bound to cellular nucleic acids (4).
The CHEMICON® Cell Invasion Assay Kit provides an efficient system for evaluating the invasion of tumor cells through a basement membrane model. The kit utilizes ECMatrixTM, a reconstituted basement membrane matrix of proteins derived from the Engelbreth Holm-Swarm (EHS) mouse tumor (5-8). We examined the kit's performance using human fibrosarcoma (HT-1080) and non-invasive fibroblasts (NIH3T3).
Test Principle
The CHEMICON® Cell Invasion Assay is performed in an Invasion Chamber, based on the Boyden chamber principle. Each kit contains 24 inserts; each insert contains an 8 μm pore size polycarbonate membrane coated with a thin layer of ECMatrixTM. The ECM layer occludes the membrane pores, blocking non-invasive cells from migrating through. Invasive cells, on the other hand, migrate through the ECM layer and cling to the bottom of the polycarbonate membrane. Invaded cells on the bottom of the insert membrane are dissociated from the membrane when incubated with Cell Detachment Buffer and subsequently lysed and detected by CyQuant GR® dye.
The ability to study cell invasion through an ECM barrier, is of vital importance for developing possible metastatic inhibitors and therapeutics. The new CHEMICON® QCM™ 24-well Invasion Assay (ECM554) provides an efficient, in vitro system for quantitative analys |