Cultrex In Vitro Angiogenesis Assay (EC Invasion), 96-well
Cultrex In Vitro Angiogenesis Assay (EC Invasion), 96-well Summary
Why Use Cultrex In Vitro Angiogenesis Assay (EC Invasion), 96-well?
The Cultrex In Vitro Angiogenesis Assay Endothelial Cell Invasion Kit was created in an effort to accelerate the screening process for compounds that influence vascular endothelial cell digestion of and invasion through a basement membrane extract (BME) layer. This kit offers a flexible, standardized, 96 well high-throughput format for quantifying endothelial invasion, in vitro, in response to chemoattractants and/or inhibiting compounds. This assay employs a simplified Boyden chamber-like design with an 8 micron polyethylene terephthalate (PET) membrane. Sulforaphane [1-isothiocyanato-(4R)-methylsulfinyl)-butane], a naturally occurring cancer chemopreventive agent, is provided as a control for inhibition of in vitro endothelial cell invasion through Cultrex BME. Detection of cell invasion is quantified using Calcein-AM, which is internalized by the cells, and converted to the acetomethylester (AM) moiety by intracellular esterases. Free Calcein fluoresces brightly, and this fluorescence may be used to quantify the number of cells that have invaded using a fluorescence plate reader.
• 5X BME Coating Solution
• Calcein AM
• Cell Invasion Chamber
• 10X Coating Buffer
• 25X Cell Wash Buffer
• 10X Cell Dissociation Solution
Note: The components for this kit may require different storage/shipping temperatures and may arrive in separate packaging.
For research use only. Not for diagnostic use.
Detection of Endothelial Cell Invasion using the Cultrex In Vitro Angiogenesis Assay. The ability of the SVEC4-10 mouse endothelial cell line, human umbilical vein endothelial cells (HUVECs), and human brain microvascular endothelial cells (HBMVECs) to cross a basement membrane extract-coated polyester filter in the absence of angiogenic growth factors (green bars), in response to angiogenic growth factors such as fetal bovine serum, VEGF, and FGF basic (dark blue bars), or in response to angiogenic growth factors in the presence of sulforaphane, an inhibitor of angiogenesis (light blue bars) was quantified using the Cultrex In Vitro Angiogenesis Assay (Endothelial Cell Invasion) Kit (Catalog # 3471-096-K).
The kit comes with both black and clear microplates. Do I need to use both?
The entire assay may be conducted in either the clear or black assay plate (provided in3455-096-01), if desired. The black assay plate provides increased sensitivity and reduced background for plate readers that read from the top. The clear assay plate provides compatibility for plate readers that read from the bottom. Some researchers have used the clear assay plate for the invasion step and the black assay plate for detection (top read).
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