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48-602MAG
Buffer Detection Kit for Magnetic Beads
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HTS112M
Sigma-AldrichChemiScreen™ Recombinant Human Glucagon Receptor Membrane Preparation
Human glucagon / GCG GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Pellicon® 3 Cassette with Ultracel® 3 kDa Membrane, C screen, 0.57 m²
Übersicht
Replacement Information
Key Spec Table
Species
Host Cells
Protein Target
Human
Chem-1
glucagon / GCG
Description
Catalogue Number
HTS112M
Brand Family
Chemicon®
Trade Name
ChemiScreen
Chemicon
Description
ChemiScreen™ Recombinant Human Glucagon Receptor Membrane Preparation
Overview
TRANSFECTION: Full-length human GCGR cDNA encoding Glucagon Receptor HOST CELLS: Chem-1, an adherent mammalian cell line with no endogenous glucagon receptor expression.
Background Information
Glucagon is a 29-amino acid peptide that stimulates glycogenolysis and gluconeogenesis in the liver to increase blood glucose. The receptor for glucagon is a class 2 (or class B) GPCR that signals through Gs to stimulate cAMP production (Mayo et al., 2003). Mice lacking the glucagon receptor have mild hypoglycemia after fasting, and exhibit hyperplasia of pancreatic α-cells (Gelling et al., 2003). Because of its role in promoting hyperglycemia, the glucagon receptor presents a potential target for treatment of diabetes. Chemicon's glucagon receptor 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 glucagon receptor interactions and its ligands. The membrane preparations exhibit a Kd of 3.09 nM for [125I]-glucagon. With 1 nM [125I]-glucagon, 5 or 10 μg/well glucagon receptor Membrane Prep yields greater than 10-fold signal-to-background ratio.
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: Membrane proteins were adjusted to 0.5 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C.
The physiologic effects of glucagon (GCG; MIM 138030) are mediated through the glucagon receptor, a member of the superfamily of receptors characterized by a 7-transmembrane domain structure and by their coupling via GTP-binding proteins (G proteins) to adenyl cyclase.[supplied by OMIM]
FUNCTION: SwissProt: P47871 # This is a receptor for glucagon which plays a central role in regulating the level of blood glucose by controlling the rate of hepatic glucose production and insulin secretion. The activity of this receptor is mediated by G proteins which activate adenylyl cyclase and also a phosphatidylinositol-calcium second messenger system. SIZE: 477 amino acids; 54009 Da SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein. DISEASE:SwissProt: P47871 # It is a candidate for certain defects in non-insulin- dependent diabetes mellitus (NIDDM). The variant in position 40 (Ser) is found in some NIDDM patients, but also in nondiabetic subjects. SIMILARITY: SwissProt: P47871 ## Belongs to the G-protein coupled receptor 2 family.
Incubation Conditions
RECOMMENDED ASSAY CONDITIONS: Membranes are mixed with radioactive ligand and unlabeled competitor 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]-glucagon (Perkin Elmer#:NEX-207) • 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 10-fold signal:background with [125I]-labeled glucagon at 1 nM
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
SPECIFICATIONS: 1 unit = 5 μg Bmax for [125I] glucagon binding: 16.7 pmol/mg protein Kd for [125I] glucagon binding: ~ 3.09 nM
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of Glucagon receptor membrane prep:
10 µg/well
5 µg/well
Signal:Background
10.1
35
Specific Binding (cpm)
33315
36252
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
Global Trade Item Number
Bestellnummer
GTIN
HTS112M
04053252321610
Documentation
ChemiScreen™ Recombinant Human Glucagon Receptor Membrane Preparation SDB
Glucagon, the counter-regulatory hormone to insulin, is secreted from pancreatic alpha cells in response to low blood glucose. To examine the role of glucagon in glucose homeostasis, mice were generated with a null mutation of the glucagon receptor (Gcgr(-/-)). These mice display lower blood glucose levels throughout the day and improved glucose tolerance but similar insulin levels compared with control animals. Gcgr(-/-) mice displayed supraphysiological glucagon levels associated with postnatal enlargement of the pancreas and hyperplasia of islets due predominantly to alpha cell, and to a lesser extent, delta cell proliferation. In addition, increased proglucagon expression and processing resulted in increased pancreatic glucogen-like peptide 1 (GLP-1) (1-37) and GLP-1 amide (1-36 amide) content and a 3- to 10-fold increase in circulating GLP-1 amide. Gcgr(-/-) mice also displayed reduced adiposity and leptin levels but normal body weight, food intake, and energy expenditure. These data indicate that glucagon is essential for maintenance of normal glycemia and postnatal regulation of islet and alpha and delta cell numbers. Furthermore, the lean phenotype of Gcgr(-/-) mice suggests glucagon action may be involved in the regulation of whole body composition.
Millipore offers a large selection of robust and reliable G-protein coupled receptor products, including Stable Cell Lines, Membrane Preparations, and Frozen Cells. See below for GPCR research tools. Weitere Informationen >>
