HTS094M
ChemiSCREEN Membrane Preparation Recombinant Human GAL1 Receptor
Human GAL1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
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200 UNITS
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200 UNITS
$884.00
About This Item
UNSPSC Code:
41106514
eCl@ss:
32161000
NACRES:
NA.84
$884.00
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biological source
human
Quality Level
recombinant
expressed in Chem-1 cells
manufacturer/tradename
ChemiScreen
Chemicon®
technique(s)
ligand binding assay: suitable (GTPγS)
radioligand binding assay (RLBA): suitable
NCBI accession no.
UniProt accession no.
shipped in
dry ice
General description
Galanin is a 29-30 amino acid peptide originally purified from intestine but later found to be abundant in the CNS. A family of 3 GPCRs, GAL1, GAL2 and GAL3, bind to galanin and mediate its biological effects. Galanin and its receptors have been implicated in pain, cognition, seizure activity, depression and drug addiction (Hokfelt, 2005). In particular, GAL1 mediates the anticonvulsant activity of galanin; mice lacking GAL1 are affected by spontaneous seizures, and have increased susceptibility to seizures induced by lithium-pilocarpine treatment and electrical perforant path stimulation (McColl et al., 2006; Mazarati et al., 2004). Chemicon′s GAL1 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 GAL1 interactions with galanin. The membrane preparations exhibit a Kd of 1.7 nM for [125I]-galanin. With 5 ug/well GAL1 Membrane Prep and 0.25 nM [125I]-galanin, a greater than 50-fold signal-to-background ratio was obtained.
Human GAL1
Application
Radioligand binding assay, and GTPγS binding.
Biochem/physiol Actions
GPCR Class: A
Protein Target: GAL1
Target Sub-Family: Galanin
Packaging
1 mL per vial
Quality
SPECIFICATIONS: 1 unit = 5 ug membrane preparation Bmax: 19.2 pmol/mg Kd: 1.7 nM
| 10 µg/well | 5 µg/well | |
|---|---|---|
| Signal:Background | 77.6 | 75.6 |
| Specific Binding (cpm) | 38414 | 37384 |
Specifications
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 Hepes, pH 7.4, 5 mM MgCl2, 1 mM CaCl2, 0.2% BSA, filtered and stored at 4°C Radioligand: [125I] galanin (Perkin Elmer # NEX333)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.
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 Hepes, pH 7.4, 5 mM MgCl2, 1 mM CaCl2, 0.2% BSA, filtered and stored at 4°C Radioligand: [125I] galanin (Perkin Elmer # NEX333)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.
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 50-fold signal:background with 125I-labeled galanin at 0.25 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.
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.
Legal Information
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
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.
Storage Class
12 - Non Combustible Liquids
wgk_germany
WGK 2
Certificates of Analysis (COA)
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Craig D McColl et al.
Neuropharmacology, 50(2), 209-218 (2005-10-26)
Galanin is a widely-distributed neuropeptide that acts as an endogenous anticonvulsant. We have recently generated a galanin receptor type 1 knockout mouse (Galr1(-/-)) that develops spontaneous seizures. Our aim here was to characterize the seizures by making electroencephalogram (EEG) recordings
Galanin and its receptors: introduction to the Third International Symposium, San Diego, California, USA, 21-22 October 2004.
Hokfelt, Tomas
Neuropeptides, 39, 125-142 (2005)
A Mazarati et al.
Neuroscience, 128(2), 431-441 (2004-09-08)
The neuropeptide galanin exhibits anticonvulsant effects in experimental epilepsy. Two galanin receptor subtypes, GalR1 and GalR2, are present in the brain. We examined the role of GalR1 in seizures by studying the susceptibility of GalR1 knockout (KO) mice to status
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