Overview

  • Product name

    Angiogenesis Assay Kit (In Vitro)
  • Detection method

    Fluorescent
  • Assay type

    Cell-based (qualitative)
  • Assay time

    6h 00m
  • Product overview

    Angiogenesis Assay Kit ab204726 provides a quick and robust method to measure the ability of endothelial cells to from three-dimensional tube-like structures in vitro in less than 18 hours. This tube formation assay provides a simple, easy to perform, qualitative tool for assessing angiogenesis.


    Angiogenesis assay protocol summary:
    - add extracellular matrix solution to empty culture plate and incubate for 1 hr at 37ºC to allow the solution to form a gel
    - plate cells onto the gel and add experimental treatment
    - incubate cells for 4-18 hrs to allow tube formation
    - remove incubation medium and wash cells / gel
    - add staining dye and incubate for 30 min
    - examine tube formation using light and fluorescence microscopy (green filter)

  • Notes

    Angiogenesis is a physiological process that occurs during wound healing and normal development which involves the growth of new blood vessels from pre-existing vessels. These blood vessels form highly branched, tree-like tubular networks that ensure efficient and simultaneous transport of gases, liquids, nutrients, signaling molecules, and circulating cells between tissues and organs. Angiogenesis is complex and highly regulated, with tight coordination of cell proliferation, differentiation, migration, matrix adhesion, and cell-to-cell signaling. Angiogenesis is regulated by several factors, most importantly growth factors such as vascular endothelial growth factors (VEGFs) and platelet-derived growth factors (PDGFs).

  • Platform

    Microplate reader

Properties

Images

  • Phase contrast (a, c, e) and fluorescent images (b, d, f) of endothelial cells in a tissue culture plate. (a, b) Endothelial cells grown without the Extracellular Matrix Gel, (c, d) Tube formation of endothelial cells grown on Extracellular Matrix gel. (e, f) endothelial cells grown on Extracellular Matrix gel treated with Suramin (10 μmol/L). Images were taken using Nikon TE2000 microscope.

  • HUVEC morphogenesis on Extracellular Matrix Gel. Cells (2 × 104) were plated per 1 cm2 well precoated with Extracellular Matrix Gel and grown for 18 hours (A) in the specific medium alone (positive control) or containing (B) PMA 10 µmol/L.

Protocols

References

This product has been referenced in:

  • Xie X & Percipalle P Elevated transforming growth factor ß signaling activation in ß-actin-knockout mouse embryonic fibroblasts enhances myofibroblast features. J Cell Physiol N/A:N/A (2018). Read more (PubMed: 29851084) »
  • Zafarnia S  et al. Nilotinib Enhances Tumor Angiogenesis and Counteracts VEGFR2 Blockade in an Orthotopic Breast Cancer Xenograft Model with Desmoplastic Response. Neoplasia 19:896-907 (2017). Read more (PubMed: 28938160) »
See all 2 Publications for this product

Customer reviews and Q&As

Answer

For Step 10.2.1, 50 uL of the thawed Extracellular Matrix Solution is added to each well of a pre-chilled (on ice) 96-well sterile cell culture plate. The gel is incubated in the plate only and not in growth media. You can leave the gel incubating for more than 1 hour, but after 1 hour the plate should be placed at 4C (not 37C) and covered to prevent evaporation or drying of the plate. If the wells are dried this might interfere with the assay.

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