Endothelial Cell Adhesion Assay
- Simple fluorimetric screening format
- Endothelial promoter and inhibitor controls
- Blocking antibodies included
Assay Overview
The recruitment of leukocytes into inflammatory tissues is regulated by the interactionbetween blood cells and endothelial cells that is preceded by integrin-mediated
cell adhesion. Endothelial-leukocyte cell adhesion plays a major role in cellular communication and regulation, and is of fundamental importance in the development and maintenance of tissues.
Expression of surface molecules on the vascular endothelium is altered at sites of pathological inflammation. Endothelial Cell Adhesion Molecules (ECAMs) are important mediators of leukocyte recruitment and adherence to the endothelium. ECAMs such as E-selectin, VCAM-1 and ICAM-1 are upregulated during inflammation, which initiates leukocyte adhesion to the endothelium, and ultimately contributes to disease progression or tissue damage. Expression of these ECAMs is mediated by proinflammatory cytokines such as Interleukin-1 beta and Tumor Necrosis Factor alpha, which propel the interaction of integrin multimers on the leukocyte cell surface and the ECAMs on the endothelial cells. Millipore’s Endothelial Cell Adhesion Assay is a two cell interactive assay to screen the adhesive properties of leukocytes or leukocyte-like cell lines in response to manipulation of activated or inactivated endothelial cells. The kit is supplied with regulators and inhibitors of endothelial cells to induce or repress expression of ECAMs, as well as other adhesion molecules. Additionally, the kit includes three monoclonal antibodies for adhesion blocking studies of ECAMs: ICAM-1, VCAM and E-selectin. These may be used to verify changes in expression level of adhesion molecules, or effects of integrins upon endothelium activation. The Endothelial Cell Adhesion Assay uses a homogeneous fluorescence detection format through a viable cell fluorescent compound,Calcein-AM and a standard fluorescent microplate reader. This allows for large scalescreening a quantitative comparison of multiple samples with the flexibilityfor testing a variety of conditions. Each kit contains sufficient reagents for the evaluation of 96 samples.
Kit Components
- 96-well Black Fluorescence Plate
- Inhibitors
- Cycloheximide
- Actinomycin D
- Proinflammatory Cytokines
- Tumor Necrosis Factor alpha
- Interleukin-1 beta
- Inhibitors
- Cycloheximide
- Actinomycin D
- Assay Buffer
- Calcein-AM Dye
- Blocking Antibodies
- ICAM-1
- VCAM
- E-selectin
- Mouse Negative Control IgG
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