HTS050M
ChemiScreen H1 Membrane Preparation
Human H1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
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About This Item
UNSPSC-Code:
41106514
eCl@ss:
32161000
NACRES:
NA.84
Empfohlene Produkte
Biologische Quelle
human
Qualitätsniveau
Rekombinant
expressed in Chem-3 cells
Verpackung
pkg of 2 mL
Hersteller/Markenname
ChemiScreen
Chemicon®
Konzentration
0.5 mg/mL
Methode(n)
ligand binding assay: suitable (GTPγS)
radioligand binding assay (RLBA): suitable
NCBI-Hinterlegungsnummer
UniProt-Hinterlegungsnummer
Versandbedingung
dry ice
Allgemeine Beschreibung
Full-length human HRH1 cDNA
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.
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.
Anwendung
Radioligand binding assay, and GTPγS binding.
Biochem./physiol. Wirkung
GPCR Class: A
Protein Target: H1
Target Sub-Family: Histamine
Qualität
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of TP membrane prep:
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
| 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
Spezifikationen
Inucbation 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
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
Physikalische Form
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.
Lagerung und Haltbarkeit
Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Rechtliche Hinweise
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Haftungsausschluss
RESEARCH USE ONLY. This product is regulated in France when intended to be used for scientific purposes, including for import and export activities (Article L 1211-1 paragraph 2 of the Public Health Code). The purchaser (i.e. enduser) is required to obtain an import authorization from the France Ministry of Research referred in the Article L1245-5-1 II. of Public Health Code. By ordering this product, you are confirming that you have obtained the proper import authorization.
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.
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.
Lagerklassenschlüssel
12 - Non Combustible Liquids
WGK
WGK 2
Flammpunkt (°F)
Not applicable
Flammpunkt (°C)
Not applicable
Analysenzertifikate (COA)
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