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HTS139M ChemiSCREEN™ Membrane Preparation Recombinant Human sst5 Somatostatin Receptor

Human sst5 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: P35346 Entrez Gene Number: NM_001053.1
HTS139M
200 units  
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Spec Table

SpeciesHost CellsProtein Target
Human Chem-1sst5
Description
Catalogue NumberHTS139M
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human sst5 Somatostatin Receptor
OverviewFull-length human SSTR5 cDNA encoding sst5
Background InformationSomatostatin (sst) is a multifunctional peptide with two biologically active forms, sst-14 and sst-28, which are synthesized in neurons throughout the brain as well as in peripheral tissues such as the pancreas and the gut (Gillies, 1997). SST exerts a diverse array of effects that include inhibition of endocrine secretion, modulation of neurotransmission, and regulation of cell proliferation by stimulating a family of five G-protein-coupled receptors. Somatostatin receptor sst5 is an inhibitory G protein-coupled receptor that exerts a strong cytostatic effect on various cell types. In mice, sst5 mediates inhibition by somatostatin of pancreatic insulin secretion and contributes to the regulation of glucose homeostasis and insulin sensitivity (Strowski et al. 2003). In addition, deficiency of sst5 leads to subtype-selective sexually dimorphic changes in the expression of both brain and pancreatic somatostatins (Ramirez et al. 2004). Millipore's sst5 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 agonists and antagonists of sst5. The membrane preparations exhibit a Kd of 1.2 nM for [125I]-Somatostatin 14. With 0.5 nM [125I]-Somatostatin 14, 5 μg/well sst5 Membrane Prep typically yields greater than 10-fold signal-to-background ratio.
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 1 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C
Quality LevelMQ100
Applications
ApplicationHuman sst5 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application NotesRadioligand binding assay and GTPγS binding.
Biological Information
Entrez Gene Number
Entrez Gene SummarySomatostatin acts at many sites to inhibit the release of many hormones and other secretory proteins. The biological effects of somatostatin are probably mediated by a family of G protein-coupled receptors that are expressed in a tissue-specific manner. SSTR5 is a member of the superfamily of receptors having seven transmembrane segments.
Gene Symbol
  • SSTR5
  • SS5R
GPCR ClassA
Host CellsChem-1
Protein Targetsst5
Target Sub-FamilySomatostatin
UniProt Number
UniProt SummaryFUNCTION: SwissProt: P35346 # Receptor for somatostatin 28 and to a lesser extent for somatostatin-14. The activity of this receptor is mediated by G proteins which inhibit adenylyl cyclase.
SIZE: 364 amino acids; 39202 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Adult pituitary gland, heart, small intestine, adrenal gland, cerebellum and fetal hypothalamus. No expression in fetal or adult kidney, liver, pancreas, uterus, spleen, lung, thyroid or ovary.
SIMILARITY: SwissProt: P35346 ## 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, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) 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]-Somatostatin 14. (Perkin Elmer#:NEX-389 )

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 somatostatin 14 at 0.5 nM.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of sst5 Receptor membrane prep.
Product Usage Statements
10 µg/well 5 µg/well
Signal:Background 15.2 15.7
Specific Binding (cpm) 25710 21425

SPECIFICATIONS: 1 unit = 5 µg membrane preparation
Bmax for [125I]-Somatostatin binding: 10.4 pmol/mg protein
Kd for [125I]-Somatostatin binding: ~1.2 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
HTS139M 04053252325359

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