merck millipore,默克密理博,17-10398,In Vitro Vascular Permeability Imaging Assay (Green)

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merck millipore,默克密理博,17-10398,In Vitro Vascular Permeability Imaging Assay (Green)
产品名称：In Vitro Vascular Permeability Imaging Assay (Green)
产品型号：17-10398

merck millipore,默克密理博,17-10398,In Vitro Vascular Permeability Imaging Assay (Green)

产品介绍

merck millipore,默克密理博,17-10398,In Vitro Vascular Permeability Imaging Assay (Green)

重要规格表

检测方法
Fluorescent

描述
产品目录编号	17-10398
品牌系列	Chemicon®
商名	Chemicon
描述	In Vitro Vascular Permeability Imaging Assay (Green)
概述	ALSO AVAILABLE! 96-well ( ECM642) & 24-well ( ECM644) Collagen Pre-coated Versions Introduction: A fundamental requirement for the physiological performance of organs is the formation of diffusion barriers that separate and maintain compartments of different structure. The endothelial cell lining of the internal vasculature defines a semi-permeable barrier between the blood and the interstitial spaces of the body. This barrier is composed of intercellular adherens, tight, and gap junction complexes, as well as desmosomes. Junction substructure components such as connexins, integrins, cadherins, catenins, occludins, desmoplakins, selectins, and platelet endothelial cell adhesion molecule-1 (PECAM-1) all act as interface regulators for paracellular permeability of ions, nutrients, therapeutic agents, and macromolecules. Endothelial cell adhesive characteristics provide strength and stability for neighboring cells and the cellular cytoskeleton by interacting with actin and myosin contractile filaments. Junctional molecules also influence cell signaling and trigger responses that are translated into cell morphology changes and physiological angiogenesis. A multitude of vasoactive cytokines, growth factors, and signal modulators react with endothelial cell substructural components to control permeability. Vascular endothelial growth factor (VEGF), interleukin-1 alpha and beta (IL-1α and IL-1β), tumor necrosis factor-alpha (TNF-α), and interferon-gamma (IFN-γ) have been shown to increase endothelial monolayer permeability. Thrombin stimulation of cytoskeletal signaling pathways has been shown to increase cell permeability. In contrast, junctional adhesion molecule (JAM) decreases permeability by initiating cell adhesion and angiopoietin-1 (Ang-1) can protect endothelial barrier function through regulation of junctional complexes. Disruptions of the barrier integrity are manifested as microvascular hyperpermeability, which is associated with many systemic disease states. Pathological angiogenic disease states include heart disease, diabetes, cancer, stroke, hypertension, arthritis, and Alzheimer’s. Increases in tissue permeability may be caused by weak, hemorrhaging vessels that become oedematous, and intensifies with irregular fluid flow through the vessels. Expanding the knowledge of endothelial junction behavior and the agents that influence that behavior will lead to new therapies for controlling endothelial permeability. EMD Millipore’s In Vitro Vascular Permeability Imaging Assay provides optimized materials and protocols to enable detailed spatial analysis of intercellular permeability in endothelial monolayers. All of the components necessary for affixing a thin film of biotinylated matrix to glass culture surfaces and staining sites of intercellular permeability are provided. In addition, compatible reagents are provided for correlating tight junctions (VE-cadherin) and nuclei with sites of permeability. This assay may be used for assessing activity of inhibitors and promoters of vascular permeability, and correlating permeability with ultrastructural features and signaling events. Test Principle: The EMD Millipore In Vitro Vascular Permeability Imaging Assay provides the reagents necessary for affixing a thin, uniform layer of biotinylated gelatin to a glass culture substrate which, in the presence of an endothelial monolayer, binds to a fluorescently labeled streptavidin only at sites of intercellular permeability. A poly-L-lysine coating is first adsorbed to the glass substratum. The substrate is then treated with a dilute glutaraldehyde solution to bi-functionally “activate” the surface for further protein binding. Subsequent incubation of the surface with biotinylated gelatin allows covalent coupling between the poly-L-lysine and gelatin via reactive aldehyde (-CHO) groups. The biotin-coated glass is now prepared for cell culture by disinfection with 70% ethanol, followed by quenching of free aldehydes with amino acid-containing growth medium. The endothelial cell of interest is seeded onto the gelatin surface and allowed to form a confluent monolayer. Treatment compounds of interest may be introduced at desired time points during the culture period. Sites of intercellular permeability are then stained with fluorescent streptavidin [19], which are microscopically visualized and may be quantified using image analysis software algorithms. The assay also provides anti-VE-cadherin and DAPI, for visualization of adherens junctions and nuclei, respectively, to allow colocalization of sites of adherens junction remodeling and increased permeability. The basic method allows potential activators or inhibitors to be investigated for their influence on the degree and sites of vascular permeability. The assay may be further combined with immunocytochemical staining for other molecules of interest to colocalize sites of permeability with signaling events.

产品信息
部件	Donkey anti-Mouse IgG, Cy3 Conjugate Secondary Poly-L-Lysine, 2X DAPI Glutaraldehyde, 16X Biotin-Gelatin Fluorescein-Streptavidin VE Cadherin Primary Antibody
检测方法	Fluorescent

应用
应用说明	The EMD Millipore In Vitro Vascular Permeability Imaging Assay kit is ideal for interrogating compounds that may disrupt or protect an endothelial monolayer. Each In Vitro Vascular Permeability Imaging Assay kit contains sufficient reagents for the coating of four 8-well glass chamber slides, although alternate substrate formats may also be utilized. The EMD Millipore In Vitro Vascular Permeability Imaging Assay is intended for research use only, not for diagnostic or therapeutic applications.

应用

应用说明

The EMD Millipore In Vitro Vascular Permeability Imaging Assay kit is ideal for interrogating compounds that may disrupt or protect an endothelial monolayer. Each In Vitro Vascular Permeability Imaging Assay kit contains sufficient reagents for the coating of four 8-well glass chamber slides, although alternate substrate formats may also be utilized. The EMD Millipore In Vitro Vascular Permeability Imaging Assay is intended for research use only, not for diagnostic or therapeutic applications.

生物信息
品种反应性	All

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	Store Poly-L-Lysine and DAPI at 2-8°C. Store Biotin-Gelatin, Glutaraldehyde, fluorescein-streptavidin, anti-VE cadherin, and Donkey anti-Mouse IgG, Cy3 conjugate at -20°C. Use within 4 months from date of receipt.