Leukocyte Transendothelial Migration Pathway Luminex Assay (CAT#: STEM-MB-0360-WXH)

Introduction

The migratory properties of white blood cells are indispens able to drive immune responses throughout the body. To ensure migration to the proper locations, the trafficking ot leukocytes is tightly regulated. The migration and extravasation of leukocytes across the endothelium that lines the vessel wall occurs in several distinct steps. The first step comprises the rolling of the leukocytes over the endothelial cells. mediated by transient weak interac tions between adhesion molecules. Subsequently, loosely at- tached leukocytes are in such close proximity of the endotheliun that they can be activated by chemotactic cytokines, presented on the apical surface of the endothelium. As a consequence,the activated leukocytes will spread and firmly adhere to the endothelium and finally migrate through the intercellular clefts between the endothelial cells to the underlying tissue.

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Principle Applications Procedure Materials Notes Related Products

Principle

During the diapedesis of leukocytes, the leukocytes bind to endothelial cell adhesion molecules (CAM) and then migrate across the vascular endothelium. A leukocyte adherent to CAMs on the endothelial cells moves forward by leading-edge protrusion and retraction of its tail. In this process, alphaL /beta2 integrin activates through Vav1, RhoA, which subsequently activates the kinase p160ROCK. ROCK activation leads to MLC phosphorylation, resulting in retraction of the actin cytoskeleton. Moreover, Leukocytes activate endothelial cell signals that stimulate endothelial cell retraction during localized dissociation of the endothelial cell junctions. ICAM-1-mediated signals activate an endothelial cell calcium flux and PKC, which are required for ICAM-1 dependent leukocyte migration. VCAM-1 is involved in the opening of the "endothelial passage" through which leukocytes can extravasate. In this regard, VCAM-1 ligation induces NADPH oxidase activation and the production of reactive oxygen species (ROS) in a Rac-mediated manner, with subsequent activation of matrix metallopoteinases and loss of VE-cadherin-mediated adhesion.

Applications

To study the regulation mechanism of leukocyte transendothelial migration signal pathway in disease
To study the effect of each virus on leukocyte transendothelial migration signaling pathway
To study the effects of drugs or therapies on leukocyte transendothelial migration signaling pathways

Procedure

• Luminex Multiplex Assay
• Enzyme-linked immunosorbent assay (ELISA)
• Flow cytometry (FACS analysis) technology

Notes

Detectable targets: ACTB, ACTN1, AFDN, ARHGAP35, ARHGAP5, BCAR1, CD99, CDC42, CDH5, CLDN20, CTNNA1, CTNNB1, CTNND1, CXCL12, CXCR4, CYBA, CYBB, ESAM, EZR, F11R, Mda-5, NFκB, RIG-1, TRAF3, GAS, IRF9, MEKK1, p38, RIP1, TRAF5, Histone-H3, IRF5, MEK3, p38MAPK, SH2, TRAF6

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