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HTS042M ChemiScreen™ BLT1 Membrane Preparation

Human BLT1 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.

ChemiScreen™ BLT1 Membrane Preparation MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: Q15722 Entrez Gene Number: NM_181657.1
HTS042M
200 units  
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Spec Table

SpeciesHost CellsProtein Target
Human Chem-1BLT1
Description
Catalogue NumberHTS042M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiScreen™ BLT1 Membrane Preparation
OverviewHuman BLT1
Background InformationBLT1 receptor is a Gi-coupled GPCR that binds primarily to the eicosanoid leukotriene B4. Macrophages, leukocytes and eosinophils express BLT1, and in vivo, BLT1 appears to mediate leukocyte migration and inflammation (reviewed in Brink et al., 2003). Chemicon's BLT1 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 BLT1 interactions with Leukotriene B4. The membrane preparations exhibit a Kd of 1.3 nM for [3H]-Leukotriene B4. With 5.0 μg/well BLT1 Membrane Prep and 1 nM [3H]-Leukotriene B4, a signal-to-background ratio of approximately 3-fold is obtained.
References
Product Information
FormatMembranes
PresentationLiquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives.

Packaging method: Membranes protein were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
Quality LevelMQ100
Applications
ApplicationHuman BLT1 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application NotesRadioligand binding assay, and GTPgammaS binding.
Biological Information
Concentration1 mL, 0.5 mg/mL per vial.
Entrez Gene Number
Gene Symbol
  • LTB4R
  • BLTR
  • BLT1
  • LTBR1
  • CMKRL1
  • P2Y7
  • GPR16
  • P2RY7
  • BLT
  • LTB4R1
GPCR ClassA
Host CellsChem-1
Protein TargetBLT1
Target Sub-FamilyLeukotriene
UniProt Number
UniProt SummaryFUNCTION: SwissProt: Q15722 # Receptor for extracellular ATP > UTP and ADP. The activity of this receptor is mediated by G proteins which activate a phosphatidylinositol-calcium second messenger system. May be the cardiac P2Y receptor involved in the regulation of cardiac muscle contraction through modulation of L-type calcium currents. Is a receptor for leukotriene B4, a potent chemoattractant involved in inflammation and immune response.
SIZE: 352 amino acids; 37557 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Expressed at highest levels in heart, skeletal muscle and at lower levels in brain and liver. High level of expression in lymphoid tissues.
PTM: Phosphorylated by GRK6 upon leukotriene B4 binding; which promotes desensitization.
SIMILARITY: SwissProt: Q15722 ## Belongs to the G-protein coupled receptor 1 family.
Incubation ConditionsRECOMMENDED ASSAY 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, a GF/C 96-well filter plate is coated with 0.33% polyethyleneimine for 30 min, then washed with 50mM HEPES, pH 7.4, 0.5% BSA. 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 Hepes, pH 7.4, 5 mM MgCl2, 1 mM CaCl2, 0.2% BSA, filtered and stored at 4°C

Radioligand: [3H] Luekotriene (Perkin Elmer# NEX378)

Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.

One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 2.5-fold signal:background or 500 cpm with 3H-labeled Leukotriene B4 at 1 nM
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
SPECIFICATIONS:
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of Motilin Receptor membrane prep:
Product Usage Statements
5 µg/well 10 µg/well
Signal:Background 75 65
Specific Binding (cpm) 46348 54457


1 unit = 5µg membrane preparation
Bmax: 5.6 pmol/mg
Kd: 0.43 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 ConditionsMaintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Packaging Information
Material Size200 units
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
HTS042M 04053252737701

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