Key Specifications Table
|Description||QCM™ Endothelial Cell Invasion Assay (24 well, fluorometric)|
|Overview||Also available: Cell Comb™ Scratch Assay! Get biochemical data from a scratch assay! Click Here
Endothelial cells (EC) invade through the basement membrane (BM) to form sprouting vessels. The invasion process consists of the secretion of matrix metalloproteases (MMP) to degrade basement membrane, the activation of endothelial cells, and the migration of EC across the basement membrane. The understanding of EC invasion is important for studying the mechanism of angiogenesis in injured tissue as well as in disease such as cancer.
Cell migration may be evaluated through several different methods, the most widely accepted of which is the Boyden Chamber assay. The Boyden Chamber system uses two-chamber system which a porous membrane provides an interface between two chambers. Cells are seeded in the upper chamber and chemoattractants placed in the lower chamber. Cells in the upper chamber migrate toward the chemoattractants by passing through the porous membrane to the lower chamber. Migratory cells are then stained and quantified.
|Materials Required but Not Delivered||1. Precision pipettes: sufficient for aliquoting cells.
2. Harvesting buffer: EDTA or trypsin-based cell detachment buffer, or Millipore’s non-mammalian cell detachment solution, Accutase™ (Cat. No. SCR005) as a gentle alternative.
3. Endothelial cells, for example: HUVECs cells (Cat. No. SCCE001)
4. Endothelium cell culture medium appropriate for subject cells, such as EGM-2 (Endothelial cell growth media-2)
5. Quenching Medium: serum-free medium, such as EBM-2 etc containing 5% BSA. Must contain divalent cations (Mg2+, Ca2+) sufficient for quenching EDTA in harvesting buffer.
6. Sterile PBS or HBSS to wash cells.
7. Distilled water
8. (Optional) Chemoattractant or pharmacological agent added to culture medium
9. Low speed centrifuge and tubes for cell harvesting.
10. CO2 incubator appropriate for subject cells.
11. Hemocytometer or other means of counting cells.
12. Trypan blue or equivalent viability stain.
13. Microplate reader with Excitation/Emission of 495/520 nm or FITC channel
14. Sterile cell culture hood
15. (Optional) Graduated ocular (calibrated), or automated method for counting stained cells on a membrane.
|Safety Information according to GHS|
|Material Size||1 kit|
|Material Package||Sufficient for 24 samples|
|QCM™ Endothelial Cell Invasion Assay (24 well, fluorometric)|