ECM211 QCM™ Endothelial Cell Invasion Assay (24 well, fluorometric)

ECM211
1 kit  Sufficient for 24 samples
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      Overview

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      Key Specifications Table

      Detection Methods
      Fluorescent
      Description
      Catalogue NumberECM211
      Trade Name
      • QCM
      • Chemicon
      DescriptionQCM™ Endothelial Cell Invasion Assay (24 well, fluorometric)
      OverviewAlso available: Cell Comb™ Scratch Assay! Get biochemical data from a scratch assay! Click Here

      Introduction
      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 Delivered1. 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.
      16. Shaker
      References
      Product Information
      Components
      • ECM211-1
      • 1. 24-well Cell Invasion Plate Assembly: (Part No. CS203020) Two 24-well plates each containing 12 ECMatrix™-coated 3 µm inserts per plate.
      • 2. 24-well Stain Extraction Plate: (Part No. 2005871) One each.
      • 3. 96-well Quantitation Plate: (Part No. CR201763) One each.
      • 4. Forceps: (Part No. 10203) One each.
      • ECM211-2
      • 1. Cell Detachment Solution: Accutase™ (Cat. No. SCR005) One 100 mL bottle.
      • 2. Calcein-AM: (Part No. CS202541) One 50 µg vial.
      Detection methodFluorescent
      Applications
      ApplicationThis QCM Endothelial Cell Invasion Assay provides an in vitro model to quickly screen factors that can regulate endothelial invasion.
      Application NotesMillipore’s QCM Endothelial Cell Invasion Assay provides an in vitro model to quickly screen factors that can regulate endothelial invasion. The assay is performed in an invasion chamber using a basement membrane protein coated on the porous insert. The level of coating and the pore size is optimized for endothelial cells so the researcher may utilize this kit to mimic physiological condition. After cell invasion has occurred, the researcher can select a staining method to quantify the number of cells that have invaded through the chamber. Millipore offers a colorimetric staining kit (ECM210) and a fluorometric kit staining reagent (ECM211) for both convenience and efficiency.
      Biological Information
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsECM211-1 kit components should be stored at 2° to 8°C up to the expiration date provided on the kit. DO NOT FREEZE.

      ECM211-2 kit components must be stored at -20°C up to the expiration date provided on the kit.
      Packaging Information
      Material Size1 kit
      Material PackageSufficient for 24 samples
      Transport Information
      Supplemental Information
      Specifications

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      Categories

      Life Science Research > Cell Analysis > Cell-based Assays > Cell Invasion Assays