merck millipore,默克密理博,ECM645,Endothelial Cell Adhesion Assay Kit

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merck millipore,默克密理博,ECM645,Endothelial Cell Adhesion Assay Kit
产品名称：Endothelial Cell Adhesion Assay Kit
产品型号：ECM645
The Endothelial Cell Adhesion Assay Kit is a versatile tool for assessing & quantifying cell to cell interaction of human endothelial cells with leukocytes, or other cell types, under a variety of con More>>

merck millipore,默克密理博,ECM645,Endothelial Cell Adhesion Assay Kit

产品介绍

merck millipore,默克密理博,ECM645,Endothelial Cell Adhesion Assay Kit

重要规格表

检测方法
Fluorescent

描述
产品目录编号	ECM645
品牌系列	Chemicon®
商名	Chemicon
描述	Endothelial Cell Adhesion Assay Kit
概述	Introduction: Endothelial cells form the inner lining of blood vessels and serve as the cellular interface between the circulating blood and the vessel wall. Endothelial cells are involved in many aspects of vascular biology, including inflammation and angiogenesis. The recruitment of leukocytes into inflammatory tissues is regulated by the interaction between 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. Atherosclerosis, ulcer, myocardial infarction, stroke, and other inflammatory processes are all related to, and dependent on this endothelial-leukocyte adhesion process. 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. The genes encoding ECAMs contain overlapping control mechanisms that allow a single signaling pathway to upregulate several genes with the potential to change the endothelial phenotype. Since ECAMs are regulated at the gene level by transcription factors, proteasome inhibitors can modulate cytokine-induced expression of ECAMs and subsequent leukocyte adhesion to the endothelium. Controlling and manipulating the endothelial-leukoctye adhesion process will create new avenues for drug therapies for vascular and inflammatory disease states. The CHEMICON® Endothelial Cell Adhesion Assay kit (ECM645) allows the researcher to test a variety of cell types that interact with activated or inactivated endothelial cell layers. Human umbilical vein endothelial cells (HUVECs) from ATCC® (CRL-1730) or Cambrex© (CC-2519) are ideal for measuring endothelial adhesion activity with the assay. The CHEMICON® Endothelial Cell Adhesion Assay kit includes a 96 well tissue culture treated fluorescence microtiter plate and reagents that allow for large-scale screening and quantitative comparison of multiple samples/cell line adhesion to an endothelial monolayer. Once endothelial cells are seeded in the tissue culture plate, they can be treated with cytokines, chemokines, transcription regulators, or selective adhesion inhibitors. The protein synthesis inhibitor Cycloheximide and the RNA synthesis inhibitor Actinomycin D are provided as controls for adhesion protein expression studies. Treatment of endothelial cells, such as human umbilical vein endothelial cells (HUVECs), with pro-inflammatory cytokines such as Interleukin-1ÃÅ¸ eta or Tumor Necrosis Factor-a can induce expression of ECAMs, as well as other adhesion molecules. The kit is supplied with both cytokines. The viable cell fluorescent compound Calcein-AM® is provided for labeling test cells, such as leukocytes (granulocytes, monocytes, lymphocytes), fibroblasts, neural stem cells etc., and quantifying their adhesion by comparing bound versus unbound cells. The kit includes three Mouse anti-human monoclonal mAbs for adhesion blocking studies of ICAM-1, VCAM, and E-selectin endothelial cell surface markers. These may be used to verify changes in expression level of adhesion molecules, or effects of integrins upon endothelium activation. This simple and high-throughput kit format allows for the monitoring of the effects of a variety of conditions and compounds on endothelial adhesion. CHEMICON® also offers a variety of antibodies to leukocyte adhesion molecules and surface antigens. CHEMICON® continues to provide numerous migration, invasion, and adhesion products including: Endothelial Cell Characterization Kit (SCR023) In Vitro Angiogenesis Assay (ECM625) Fibrin In Vitro Angiogenesis Assay (ECM630) Blood Vessel Staining Kit (ECM590) Alpha Integrin-Mediated Cell Adhesion Colorimetric Array Kit (ECM530) Beta Integrin-Mediated Cell Adhesion Colorimetric Array Kit (ECM531) Alpha/Beta Integrin-Mediated Cell Adhesion Colorimetric Array Combo Kit (ECM532) Alpha Integrin-Mediated Cell Adhesion Fluorimetric Array (ECM533) Beta Integrin-Mediated Cell Adhesion Fluorimetric Array (ECM534) Alpha/Beta Integrin-Mediated Cell Adhesion Array Combo Fluorimetric Kit (ECM535) ECM Cell Adhesion Colorimetric Array Kit (ECM540) ECM Cell Adhesion Fluorimetric Array Kit (ECM545) QCM™ 8μm 96-well Chemotaxis Cell Migration Assay (ECM510) QCM™ 8μm 24-well Chemotaxis Cell Migration Colorimetric Assay (ECM508) QCM™ 8μm 24-well Chemotaxis Cell Migration Fluorimetric Assay (ECM509) QCM™ 5μm 96-well Chemotaxis Cell Migration Assay (ECM512) QCM™ 3μm 96-well Chemotaxis Cell Migration Assay (ECM515) QCM™ 96-well ECMatrixTM Cell Invasion Assay (ECM555) QCM™ 96-well Collagen-based Cell Invasion Assay (ECM 556) Test Principle: The CHEMICON® Endothelial Cell Adhesion Assay utilizes a homogenous fluorescence detection format, allowing for large-scale screening and quantitative comparison of multiple samples with the flexibility for testing a variety of conditions. First, endothelial cells are seeded in the 96 well plate where they grow to confluency. Prior to activation, the endothelium can be treated with the protein synthesis inhibitor, (Cycloheximide) or the RNA synthesis inhibitor (Actinomycin D) to control endothelial adhesion protein expression. Subsequently, the endothelium is activated with one or two cytokines (TNF alpha and Interleukin-1 beta) provided. Upon activation, endothelial cells can be used for adhesion blocking studies with functional blocking mAbs to endothelial cell adhesion markers (e.g. E-selectin, ICAM-1, VCAM). Next, test cells are labeled with the Calcein AM® green fluorescence viability cell marker and added to the endothelium on the plate. After a brief incubation, unbound cells are washed away while bound cells remain attached to the endothelium. Finally, the plate is read on a fluorescence plate reader. Relative test cell adhesion is directly related to the fluorescence of the wells. See the diagram on the next page.

产品信息
部件	Mouse Negative Control IgG: (Chemicon Cat. No. PP54-100UG) One vial - 100 μL of solution. 96-Well Endothelial Cell Tissue Culture Fluorescence Plate: (Part No.: 2004174) One 96-well tissue culture-treated black fluorescence plate with lid. Cycloheximide: (Part No. 2004165) One vial -100 μL of solution. Actinomycin D: (Part No. 2004166) One vial - 50 μg of crystals. Tumor Necrosis Factor alpha: (Part No. 2004167) (Chemicon Cat. No. GF023) One vial - 20 μL of solution. Interleukin-1 beta: (Part No. 2004168) (Chemicon Cat. No. IL038) One vial - 12 μL of solution. 1X Assay Buffer: (Part No. 2004320) One bottle - 100 mL of solution. Calcein AM Dye-: (Part No. 2004169) One vial - 50 μL of solution, ready to use. Mouse anti-Human ICAM-1 Monoclonal Antibody: (Part No. 2004181) (Chemicon Cat. No. MAB1379) One vial - 30 μL of solution. Mouse anti-Human VCAM Monoclonal Antibody: (Part No. 2004182) (Chemicon Cat. No. MAB2144) One vial - 30 μg in solution. Mouse anti-Human E-selectin Monoclonal Antibody: (Part No. 2004183) One vial - 37.5 μL of solution.
检测方法	Fluorescent

产品信息

部件

Mouse Negative Control IgG: (Chemicon Cat. No. PP54-100UG) One vial - 100 μL of solution.
96-Well Endothelial Cell Tissue Culture Fluorescence Plate: (Part No.: 2004174) One 96-well tissue culture-treated black fluorescence plate with lid.
Cycloheximide: (Part No. 2004165) One vial -100 μL of solution.
Actinomycin D: (Part No. 2004166) One vial - 50 μg of crystals.
Tumor Necrosis Factor alpha: (Part No. 2004167) (Chemicon Cat. No. GF023) One vial - 20 μL of solution.
Interleukin-1 beta: (Part No. 2004168) (Chemicon Cat. No. IL038) One vial - 12 μL of solution.
1X Assay Buffer: (Part No. 2004320) One bottle - 100 mL of solution.
Calcein AM Dye-: (Part No. 2004169) One vial - 50 μL of solution, ready to use.
Mouse anti-Human ICAM-1 Monoclonal Antibody: (Part No. 2004181) (Chemicon Cat. No. MAB1379) One vial - 30 μL of solution.
Mouse anti-Human VCAM Monoclonal Antibody: (Part No. 2004182) (Chemicon Cat. No. MAB2144) One vial - 30 μg in solution.
Mouse anti-Human E-selectin Monoclonal Antibody: (Part No. 2004183) One vial - 37.5 μL of solution.

检测方法

Fluorescent

应用
应用	The Endothelial Cell Adhesion Assay Kit is a versatile tool for assessing & quantifying cell to cell interaction of human endothelial cells with leukocytes, or other cell types, under a variety of conditions.
应用说明	The CHEMICON® Endothelial Cell Adhesion Assay Kit is a versatile tool for assessing and quantifying cell to cell interaction of human endothelial cells with leukocytes, or other cell types, under a variety of conditions. The kit is ideal for the assessment of specific cell surface integrins and surface receptors on human endothelial cells, monitoring in vitro cell differentiation, assessing endothelial adhesion blocking, or screening potential cell adhesion promoters/inhibitors. A variety of leukocytes or leukocyte-like cell lines can be tested in response to manipulation of the endothelial cell behavior. Each CHEMICON® Endothelial Cell Adhesion Assay Kit contains sufficient reagents for the evaluation of 96 samples. The CHEMICON® Endothelial Cell Adhesion Assay Kit is intended for research use only, not for diagnostic or therapeutic applications.

应用

The Endothelial Cell Adhesion Assay Kit is a versatile tool for assessing & quantifying cell to cell interaction of human endothelial cells with leukocytes, or other cell types, under a variety of conditions.

应用说明

The CHEMICON® Endothelial Cell Adhesion Assay Kit is a versatile tool for assessing and quantifying cell to cell interaction of human endothelial cells with leukocytes, or other cell types, under a variety of conditions. The kit is ideal for the assessment of specific cell surface integrins and surface receptors on human endothelial cells, monitoring in vitro cell differentiation, assessing endothelial adhesion blocking, or screening potential cell adhesion promoters/inhibitors. A variety of leukocytes or leukocyte-like cell lines can be tested in response to manipulation of the endothelial cell behavior. Each CHEMICON® Endothelial Cell Adhesion Assay Kit contains sufficient reagents for the evaluation of 96 samples.

The CHEMICON® Endothelial Cell Adhesion Assay Kit is intended for research use only, not for diagnostic or therapeutic applications.

生物信息
品种反应性	Human

产品使用声明
使用声明	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.

储存和货运信息
存储条件	The Assay Buffer and the Endothelial Cell Culture Plate can be stored at 2° to 8°C up to their expiration dates. Store the plate in its foil pouch. If not using the entire plate, seal any used wells with a plate sealer to protect the unused wells. Ensure that the desiccant remains in the pouch, and that the pouch is securely closed. Store the remaining kit components at -20°C. Prepare undiluted aliquots for the Cycloheximide, TNF alpha, and IL-1 beta. Avoid repeated freeze-thaw cycles. Note: Actinomycin D adsorbs to plastic and glass upon standing in solution; however, frozen aliquots of the stock vial are expected to be stable for at least 30 days at -20°C. Precautions: * No data is available on the biological toxicity of Calcein AM- dye. This chemical is membrane permeable and, as such, should be treated as a potential mutagen, which may cause cancer and heritable genetic damage. Handle with caution. DMSO stock solution should be handled with special caution as DMSO can facilitate the entry of organic molecules into tissues. * Cycloheximide is highly toxic and should be handled with care. Use protective wear when handling. This chemical is membrane permeable and, as such, should be treated as a potential mutagen, which may cause cancer and heritable genetic damage. * Actinomycin D is highly toxic, carcinogenic, teratogenic and should be handled with care. Use protective wear when handling. This chemical is membrane permeable and, as such, should be treated as a potential mutagen, which may cause cancer and heritable genetic damage. DMSO stock solution should be handled with special caution as DMSO can facilitate the entry of organic molecules into tissues.

储存和货运信息

存储条件

The Assay Buffer and the Endothelial Cell Culture Plate can be stored at 2° to 8°C up to their expiration dates. Store the plate in its foil pouch. If not using the entire plate, seal any used wells with a plate sealer to protect the unused wells. Ensure that the desiccant remains in the pouch, and that the pouch is securely closed.

Store the remaining kit components at -20°C. Prepare undiluted aliquots for the Cycloheximide, TNF alpha, and IL-1 beta. Avoid repeated freeze-thaw cycles.

Note: Actinomycin D adsorbs to plastic and glass upon standing in solution; however, frozen aliquots of the stock vial are expected to be stable for at least 30 days at -20°C.

Precautions:

* No data is available on the biological toxicity of Calcein AM- dye. This chemical is membrane permeable and, as such, should be treated as a potential mutagen, which may cause cancer and heritable genetic damage. Handle with caution. DMSO stock solution should be handled with special caution as DMSO can facilitate the entry of organic molecules into tissues.

* Cycloheximide is highly toxic and should be handled with care. Use protective wear when handling. This chemical is membrane permeable and, as such, should be treated as a potential mutagen, which may cause cancer and heritable genetic damage.

* Actinomycin D is highly toxic, carcinogenic, teratogenic and should be handled with care. Use protective wear when handling. This chemical is membrane permeable and, as such, should be treated as a potential mutagen, which may cause cancer and heritable genetic damage. DMSO stock solution should be handled with special caution as DMSO can facilitate the entry of organic molecules into tissues.