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QCM 3 μm Endothelial Cell Migration Assay - Fibronectin, Fluorometric

QCM

Introduction
Angiogenesis is a fundamental process involving the growth of new blood vessels from pre-existing vessels. It is important in development and wound healing, as well as pathologic diseases such as diabetic retinopathy and cancer. During angiogenesis, endothelial cells need to move out of existing vessels, migrate into new areas, proliferate and assemble into new capillaries. The migration of endothelial cells is regulated by many angiogenic factors and anti-angiogenic factors. It is critical for researchers to understand the mechanisms of endothelial cell migration.

Millipore's 3 μm QCM™ Endothelial Cell Migration Assay – Fibronectin, Fluorometric provides a quick and efficient system to study the ability of compounds to induce or inhibit endothelial cell migration. This assay also allows screening of pharmacological agents, evaluation of integrins or other adhesion receptors responsible for endothelial cell migration, analysis of gene function in transfected cells, and determination of ECM protein involvement in cell movement.

This versatile assay permits counting of individual migratory cells, and, more importantly, allows quantitative analysis by optical density (OD) using a standard microplate reader. This convenient assay allows large scale screening and quantitative comparison of multiple samples and includes individual migration controls for each sample.

The Millipore QCM™ 3 μm Endothelial Cell Migration Assay – Fibronectin, Fluorometric is ideal for the study of endothelial cell migration in response to an angiogenic stimulus. The quantitative nature of this assay is especially useful for large scale screening of pharmacologic agents. BSA-coated control chambers provide an appropriate migration control. The 3 μm pore size in this assay is optimal for endothelial cells such as HUVEC, but not sufficient for fibroblast migration. The Millipore QCM™ 3 μm Endothelial Cell Migration Assay – Fibronectin, Fluorometric assay is intended for research use only; not for diagnostic applications.
Each kit provides sufficient materials for the evaluation of 12 samples.

Vertebrates

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.

Cell Migration Assays

• 1. Fibronectin Test Plate: One 24-well culture plate, containing 12 human FN-coated Boyden chambers, sufficient for the evaluation of 12 test samples.
• 2. BSA Control Plate: One 24-well culture plate, containing 12 BSA-coated Boyden chambers, sufficient for the evaluation of 12 controls.
• 3. 4X Cell Lysis Buffer - One 16 mL bottle
• 4. CyQuant GR Dye®: One vial 75 μL
• 5. 24-Well Cell Culture Tray
• 6. 96-Well Stain Quantitation Plate
• 7. Forceps: 1 pair
• 8. Accutase (cell detachment solution): One 100 mL bottle

• The ECM201-1 kit components should be stored at 2° to 8°C up until the expiration date provided on the labels. DO NOT FREEZE.
• The accompanying 100ml bottle of Accutase also included in this kit should be stored at -20°C.

• Colorimetric
• Fluorescent

Sufficient for 12 assays

1. Endothelial cells (for example: HUVEC cells), and cell culture medium (for example: EGM-2).
2. Serum-free medium, such as serum-free EGM-2 containing 0.5% BSA.
3. Sterile PBS or HBSS to wash cells.
4. Distilled water
5. (Optional) Chemoattractant or pharmacological agent for addition to culture medium.
6. Low speed centrifuge and tubes for cell harvesting
7. CO₂ incubator appropriate for subject cells
8. Hemocytometer or other means of counting cells.
9. Trypan blue or equivalent viability stain
10. Fluorescence plate reader.