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ChemiScreen™ Membrane Prep Human Recombinant CysLT1

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货号：HTS061M

品牌：Millipore 密理博

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Replacement Information

重要规格表

Species Host Cells Protein Target
Human Chem-1 CysLT1
Description
Catalogue Number HTS061M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
Description ChemiSCREEN™ Membrane Preparation Recombinant Human CysLT1 Cysteinyl Leukotriene Receptor
Overview Full-length human CYSLTR1 cDNA
Background Information The cysteinyl leukotrienes, leukotriene C4, leukotriene D4 and leukotriene E4, are arachidonic acid derivatives modified by glutathione, Cys-Gly or Cys. Activated mast cells release cysteinyl leukotrienes, which cause smooth muscle contraction, airway constriction, and vascular permeability. The biological effects of the cysteinyl leukotrienes are mediated by two GPCRs, CysLT1 and CysLT2. A CysLT1 selective antagonist, montelukast, is used clinically in the treatment of asthma (Brink et al., 2003; Evans, 2002). Chemicon's CysLT1 membrane preparations are crude membrane preparations made from our proprietary stable recombinant cell lines to ensure high-level of GPCR surface expression; thus, they are ideal HTS tools for screening of antagonists of CysLT1 interactions and its ligands. The membrane preparations exhibit a Kd of 0.97nM for [33H]-Leukotriene D4 (LTD4). With 10.0ug/well CysLT1 membrane prep and 1.5 nM [33H]-Leukotriene D4, a greater than 3-fold signal-to-background ratio was obtained.
References
Product Information
Format Membranes
Presentation One package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 3-fold signal:background with 3H-labeled LTD4 at 1.5 nM.

Liquid in packaging buffer: 50 mM Tris, pH 7.4, 10% glycerol with no preservatives.
Packaging method: Membranes protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80oC.
Applications
Application Human CysLT1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application Notes Radioligand binding assay and GTPγS binding
Biological Information
Entrez Gene Number
Entrez Gene Summary The cysteinyl leukotrienes LTC4, LTD4, and LTE4 are important mediators of human bronchial asthma. Pharmacologic studies have determined that cysteinyl leukotrienes activate at least 2 receptors, the protein encoded by this gene and CYSLTR2. This encoded receptor is a member of the superfamily of G protein-coupled receptors. Activation of this receptor by LTD4 results in contraction and proliferation of smooth muscle, oedema, eosinophil migration and damage to the mucus layer in the lung.
Gene Symbol
  • CYSLTR1
  • CysLTR1
  • CYSLTR
  • CysLT(1)
  • MGC46139
  • HMTMF81
  • CYSLT1
  • CYSLT1R
  • HG55
  • CysLT1
GPCR Class A
Host Cells Chem-1
Protein Target CysLT1
Target Sub-Family Leukotriene
UniProt Number
UniProt Summary FUNCTION: SwissProt: Q9Y271 # Receptor for cysteinyl leukotrienes mediating bronchoconstriction of individuals with and without asthma. Stimulation by LTD4 results in the contraction and proliferation of smooth muscle, edema, eosinophil migration and damage to the mucus layer in the lung. This response is mediated via a G-protein that activates a phosphatidylinositol-calcium second messenger system. The rank order of affinities for the leukotrienes is LTD4 > LTE4 = LTC4 > LTB4.
SIZE: 337 amino acids; 38541 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Widely expressed, with highest levels in spleen and peripheral blood leukocytes. Lower expression in several tissues, such as lung (mostly in smooth muscle bundles and alveolar macrophages), placenta, small intestine, pancreas, colon and heart.
SIMILARITY: SwissProt: Q9Y271 ## Belongs to the G-protein coupled receptor 1 family.
MISCELLANEOUS: Selective antagonists, such as montelukast (Singulair), zafirlukast (Accolate) and pranlukast (Onon), are used in the treatment of the asthma crisis.
Incubation Conditions Membranes are mixed with radioactive ligand and unlabeled competitor (see Figures 1 and 2 for concentrations tested) in binding buffer in a nonbinding 96-well plate, and incubated for 1-2 h. Prior to filtration, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) is coated with 0.33% polyethyleneimine for 30 min, then washed with 50 mM Tris, pH 7.4. Binding reaction is transferred to the filter plate, and washed 3 times (1 mL per well per wash) with Wash Buffer. The plate is dried and counted.

Binding buffer: 50 mM Tris, pH7.4, 20 mM CaCl2, 25 mM MgCl2, 10 mM Glycine, 10 mM L-Cysteine, filtered and stored at 4°C
Radioligand: [3H] LTD4 (Perkin Elmer#: NET-101)
Wash Buffer: 50 mM Tris, pH 7.4, filtered and stored at 4°C.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
1 Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of CysLT Receptor membrane prep.
Product Usage Statements
10 µg/well
Signal:Background 4.62
Specific Binding (cpm) 1451.4
SPECIFICATIONS: 1 unit = 10 µg membrane preparation
Bmax: 1.53 pmol/mg
K d: 0.97 nM Usage Statement
  • 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.
Storage and Shipping Information
Storage Conditions Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Packaging Information
Material Size 200 units
Transport Information
Supplemental Information
Specifications

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