人E选择素E-selectin试剂盒
I. INTRODUCTION
E-selectin is expressed on inflamed endothelial cells in response to treatment with inflammatory cytokines. Intravital microscopic experiments have shown that its function in mediating leukocyte rolling is largely redundant with that of P-selectin. Consequently, E-selectin deficient mice have only a subtle defect in leukocyte rolling as shown by much faster rolling velocities. In addition to mediating leukocyte rolling, E-selectin participates in the conversion of rolling to firm adhesion. E-selectin deficient mice have a reduced number of firmly adherent leukocytes in response to local chemoattractant or cytokine stimulation. This defect may be related to the more rapid rolling velocities in the absence of E-selectin. E-selectin is expressed in skin microvessels under baseline conditions and there is some evidence that E-selectin is of particular importance in skin inflammation, because it supports the recruitment ofskin-specific T lymphocytes. The ligands for E-selectin that is responsible for the rolling interaction are unknown. Two candidate ligands, PSGL-1 and E-selectin ligand-1 （ESL-1） have not been shown to be required for E-selectin mediated leukocyte rolling under any condition. It is not clear whether the physiological ligand for E-selectin is a glycoprotein. Some glycolipids can support E-selectin dependent rolling in vitro E-selectin mediates much slower rolling than P-selectin. Dependent on the level ofexpression of E-selectin, rolling velocities range between less than 5 µm/s and about 15 µm/s Since the dissociation rate or off-rate of E-selectin is very similar to that of P-selectin.
人E选择素E-selectin试剂盒
II. REAGENTS
Materials provided with the kits :
. 1、Coated Microtitration Strips 96 wells.
2、Standard 20, 40, 80, 160 and 320ng/ml ，1 set.
3、Enzyme Conjugate Solution, 12 ml.
4、Substrate A, 6 ml.
5、Substrate B, 6 ml.
6、Stopping Solution （1N HCl）, 6 ml.
7、Rinsing buffer,60 ml（ x 20）.
8、dilution,15ml（x5）
Materials required but not provided :
Precision pipettes: 0.10, 0.20, and 1.0 ml.
Disposable pipette tips.
Distilled water.
Vortex mixer or equivalent.
Absorbent paper or paper towels.
Microtiter plate shaker
Graph paper.
A microtiter plate reader with a bandwidth of 10nm or less and an optical density range of 0-2 OD or greater at 450 nm .
