HTS201M

ChemiSCREEN Membrane Preparation Recombinant Human GPR109A Receptor

Human GPR109A GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

• Código UNSPSC: 41123201
• eCl@ss: 32161000

Propiedades

• origen biológico: human
• recombinante: expressed in Chem-4 cells
• fabricante / nombre comercial: ChemiScreen; Chemicon^®; Millipore
• técnicas: ligand binding assay: suitable (GTPγS); radioligand binding assay (RLBA): suitable
• Nº de acceso NCBI: NM_177551.3
• Nº de acceso UniProt: Q8TDS4
• Condiciones de envío: dry ice

Descripción

Descripción general

Full-length human GPR109A cDNA

Nicotinic acid (niacin), a vitamin of the B complex, is used clinically in high doses to decrease total plasma levels of cholesterol. Interestingly, the total cholesterol levels and low-density lipoprotein (LDL) concentrations decrease, while the high-density lipoprotein (HDL) concentrations increase with nicotinic acid treatment. The cholesterol-lowering effects of nicotinic acid result from inhibition of lipolysis in adipose tissue, which decreases plasma levels of free fatty acid (FFA) (Altschul et al., 1955; Carlson, 1963). In a study of nicotinic acid and myocardial infarction in the Coronary Drug Project (1966-1975), patients receiving 3 g/day nicotinic acid exhibited reduced rates of myocardial infarction (Coronary Drug Project Research Group, 1975). However, an unwanted effect of high doses of nicotinic acid is vasodilatation, occurring mainly in the upper body and face, known as flushing. Recently two receptors for nicotinic acid have been identified as class A G protein-coupled receptors that couple to Gi to inhibit accumulation of cAMP (Offermans, 2006). GPR109A (also known as HM74A in humans and PUMA-G in mice) is a high affinity receptor for nicotinic acid, whereas GPR109B (also known as HM74) is a low affinity receptor for nicotinic acid that is found in humans but not rodents (Wise et al., 2003). GPCR109A is found primarily in adipose tissue and immune cells. Millipore′s GPR109A 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 GPR109A interactions with its ligands. The membrane preparations exhibit EC50s of 3.65 μM for nicotinic acid in a GTPγS binding assay.

Aplicación

Radioligand binding assay and GTPγS binding.

Acciones bioquímicas o fisiológicas

GPCR Class: A

Protein Target: GPR109A

Target Sub-Family: Nicotinic Acid

Calidad

SPECIFICATIONS: 1 unit = 5 μg
EC50 in GTPγS binding assay by Nicotinic Acid: ~ 3.65 μM

Especificaciones

Inucbation Conditions
ASSAY CONDITIONS: Membranes are permeabilized by addition of saponin to an equal concentration by mass, then mixed with [³⁵S]-GTPγS (final concentration of 0.3 nM) in 20 mM HEPES, pH 7.4/100 mM NaCl/10 mM MgCl2/0.5 μM GDP in a nonbinding 96-well plate. Unlabeled nicotinic acid was added to the final concentration indicated in Figure 1 (final volume 100 μL), and incubated for 30 min at 30°C. The binding reaction is transferred to a GF/B filter plate (Millipore MAHF B1H) previously prewetted with water, and washed 3 times (1 mL per well per wash) with cold 10 mM sodium phosphate, pH 7.4. The plate is dried and counted.

One vial contains enough membranes for at least 200 assays (units), where one unit is the amount of membrane that will yield greater than 1000 cpm specific nicotinic acid-stimulated [³⁵S]-GTPγS binding.

The GPR109A membrane preparation is expected to be functional in a radioligand binding assay; however, the end user will need to determine the optimal radiolabeled ligand for use with this product.

Forma física

Liquid 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

Almacenamiento y estabilidad

Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.

Información legal

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Cláusula de descargo de responsabilidad

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.

Información de seguridad

• Código de clase de almacenamiento: 12 - Non Combustible Liquids
• Clase de riesgo para el agua (WGK): WGK 2
• Punto de inflamabilidad (°F): Not applicable
• Punto de inflamabilidad (°C): Not applicable