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ChemiScreen™ Membrane Prep Hu Recombinant FPR1

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货号：HTS018M

品牌：Millipore 密理博

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Replacement Information

重要规格表

Species Host Cells Protein Target
Human Chem-2 FPR1
Description
Catalogue Number HTS018M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
Description ChemiSCREEN™ FPR1 Membrane Preparation
Overview Human FPR1
Background Information FPR1 is a Gi-coupled GPCR that binds and is activated by short, N-formyl peptides, such as N-Formyl-Met-Leu-Phe (fMLF). N-formyl peptides are typically derived from bacteria and mediate host recruitment of leukocytes, which highly express FPR1, to sites of bacterial infection (Gao et al., 1999). In addition, FPR1 is expressed in the vasculature, secretory epithelial cells, neurons and other tissues (Becker et al., 1998). Other ligands, such as mitochondrially encoded formyl peptides, amyloid proteins, and virus-derived peptides, have also been found to activate FPR1 and/or related GPCRs and may be important in the pathophysiology of amyloidoses, Alzheimer's disease and HIV (Le et al., 2002). A synthetic hexapeptide ligand, WKYMVm, has also been demonstrated to bind to FPR1 (Le et al., 1999). Chemicon's 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 FPR1 and its ligands. The membrane preparations exhibit a Kd of 0.39 nM for [125I]-WKYMVm. With 5 μg/well FPR1 Membrane Prep and 0.5 nM [125I]-WKYMVm, a greater than 20-fold signal-to-background is obtained; with 1.25 and 2.5 μg/well FPR1 Membrane Prep, greater than 9- and 15-fold signal-to-background ratios are obtained, respectively.
References
Product Information
Format Membranes
Presentation Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives.Packaging method: Membranes protein were adjusted to 0.5 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C.
Applications
Application Human FPR1 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application Notes Radioligand binding assay, and GTPgammaS binding.
Biological Information
Concentration 0.5 mg/mL
Entrez Gene Number
Gene Symbol
  • FPR1
  • FPR
  • FMLP
GPCR Class A
Host Cells Chem-2
Protein Target FPR1
Target Sub-Family N-formylpeptide
UniProt Number
UniProt Summary FUNCTION: SwissProt: P21462 # High affinity receptor for N-formyl-methionyl peptides, which are powerful neutrophils chemotactic factors. Binding of FMLP to the receptor causes activation of neutrophils. This response is mediated via a G-protein that activates a phosphatidylinositol-calcium second messenger system.
SIZE: 350 amino acids; 38401 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Neutrophils.
PTM: Phosphorylated; which is necessary for desensitization.
SIMILARITY: SwissProt: P21462 ## Belongs to the G-protein coupled receptor 1 family.
Incubation Conditions RECOMMENDED 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: [125I] WKYMVm (Perkin Elmer# NEX386)

Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl, 0.1% BSA, filtered and stored at 4°C. One vial contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 10-fold signal:background with 125I-labeled WKYMVm at 0.5 nM
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information




SPECIFICATIONS:Signal:background and specific binding for FPR1 membrane preparation.
Product Usage Statements
5 µg/well
Signal:Background 17.9
Specific Binding (cpm) 17837



Bmax: 5.65 pmol/mg
Kd: 0.39 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 assays
Transport Information
Supplemental Information
Specifications

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