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ChemiScreen™ Membrane Prep Hu Recombinant H1

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货号：HTS050M

品牌：Millipore 密理博

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Replacement Information

重要规格表

Species Host Cells Protein Target
Human Chem-3 H1
Description
Catalogue Number HTS050M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
Description ChemiScreen™ H1 Membrane Preparation
Overview Full-length human HRH1 cDNA
Background Information Histamine exerts its biological effects through a family of four GPCRs. Stimulation of the H1 histamine receptor causes smooth muscle contraction, induces vascular permeability, and stimulates catecholamine release from the adrenal medulla. H1 is also widely expressed in brain, and activation of H1 excites several types of neurons by blocking potassium conductance (Hill et al., 1997).

Chemicon's H1 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 H1 interactions and its ligands. The membrane preparations exhibit a Kd of 4.5 nM for [3H]-Pyrilamine. With 3 nM [3H]-Pyrilamine, 5 mg/well and 10 mg/well H1 Membrane Prep typically yield greater than 3 and 6-fold signal-to-background ratios, 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 H1 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
Concentration 2 mL, 0.5 mg/mL
Entrez Gene Number
Entrez Gene Summary Histamine is a ubiquitous messenger molecule released from mast cells, enterochromaffin-like cells, and neurons. Its various actions are mediated by histamine receptors H1, H2, H3 and H4. This gene was thought to be intronless until recently. The protein encoded by this gene is an integral membrane protein and belongs to the G protein-coupled receptor superfamily. It mediates the contraction of smooth muscles, the increase in capillary permeability due to contraction of terminal venules, the release of catecholamine from adrenal medulla, and neurotransmission in the central nervous system. Multiple alternatively spliced variants, encoding the same protein, have been identified.
Gene Symbol
  • HRH1
  • H1-R
  • hisH1
GPCR Class A
Host Cells Chem-3
Protein Target H1
Target Sub-Family Histamine
UniProt Number
UniProt Summary FUNCTION: SwissProt: P35367 # In peripheral tissues, the H1 subclass of histamine receptors mediates the contraction of smooth muscles, increase in capillary permeability due to contraction of terminal venules, and catecholamine release from adrenal medulla, as well as mediating neurotransmission in the central nervous system.
SIZE: 487 amino acids; 55784 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
PTM: Potential sites of phosphorylation in the third cytoplasmic loop may play an important role in regulating signal transduction through the receptor molecule.
SIMILARITY: SwissProt: P35367 ## Belongs to the G-protein coupled receptor 1 family.
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, a GF/C 96-well filter plate is coated with 0.33% polyethyleneimine for 30 min, then washed with 50 mM 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 Tris, pH 7.4, 10 mM MgCl2, 1 mM EDTA, filtered and stored at 4°C

Radioligand: [3H] Pyrilamine (Perkin Elmer#:NET594250UC )

Wash Buffer: 50 mM Tris, pH 7.4 filtered and stored at 4°C.

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 Pyrilamine at 3 nM
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information

Signal:background and specific binding values obtained in a competition binding assay with varying amounts of TP membrane prep:
Product Usage Statements
10 µg/well
Signal:Background 6.6
Specific Binding (cpm) 925



SPECIFICATIONS: 1 unit = 10 µg for most lots. Specific activity (units/µg) may vary between lots.
Bmax: 8.3 pmol/mg
Kd: 4.5 nM
Note: Lot number MM1357015 contains 2 mL at 2 mg/mL, for which 1 unit = 20 µg 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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