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HTS186M

Sigma-Aldrich

ChemiSCREEN Membrane Preparation Recombinant Human GAL2 Galanin Receptor

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About This Item

UNSPSC Code:
41123201
eCl@ss:
32161000
NACRES:
NA.45

description

GPCR Class A
protein target GAL2

Quality Level

technique(s)

radioligand binding assay (RLBA): suitable

UniProt accession no.

Gene Information

human ... GALR2(8811)

General description

FUNCTION: SwissProt: O43603 # Receptor for the hormone galanin and for GALP. The activity of this receptor is mediated by G proteins that activate the phospholipase C/protein kinase C pathway (via Gq) and that inhibit adenylyl cyclase (via Gi).
SIZE: 387 amino acids; 41700 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Expressed abundantly within the central nervous system in both hypothalamus and hippocampus. In peripheral tissues, the strongest expression was observed in heart, kidney, liver, and small intestine.
SIMILARITY: SwissProt: O43603 ## Belongs to the G-protein coupled receptor 1 family.

Application

Radioligand binding assay and GTPγS binding.

Physical form

One package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 20-fold signal:background with 125I-labeled galanin at 0.5 nM.

Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
Packaging method: Membranes protein were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.

Storage and Stability

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

Analysis Note

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. 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, filtered and stored at 4°C
Radioligand: [125I] galanin (Perkin Elmer # NEX333)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl, filtered and stored at 4°C.

Disclaimer

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.

Certificates of Analysis (COA)

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