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Die folgenden MAPmates™ sollten nicht zusammen analysiert werden: -MAPmates™, die einen unterschiedlichen Assaypuffer erfordern. -Phosphospezifische und MAPmate™ Gesamtkombinationen wie Gesamt-GSK3β und Gesamt-GSK3β (Ser 9). -PanTyr und locusspezifische MAPmates™, z.B. Phospho-EGF-Rezeptor und Phospho-STAT1 (Tyr701). -Mehr als 1 Phospho-MAPmate™ für ein einziges Target (Akt, STAT3). -GAPDH und β-Tubulin können nicht mit Kits oder MAPmates™, die panTyr enthalten, analysiert werden.
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48-602MAG
Buffer Detection Kit for Magnetic Beads
1 Kit
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HTS186M
Sigma-AldrichChemiSCREEN™ Membrane Preparation Recombinant Human GAL2 Galanin Receptor
Human GAL2 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
ChemiSCREEN™ Membrane Preparation Recombinant Human GAL2 Galanin Receptor
Overview
Full-length human GALR2 cDNA encoding GAL2
Background Information
Galanin is a 29-30 amino acid peptide originally purified from intestine, but later found to be abundant in the CNS. It is widely distributed in tissues such as the brain, spinal cord and gut, and can regulate numerous processes including feeding, nociception, nerve regeneration, memory, neuroendocrine release, and gut secretion and contractility. Galanin elicits its physiological effects through the stimulation of at least three G protein-coupled receptors (Branchek et al. 2000). GAL2 receptor couples predominantly to the activation of phospholipase C. It plays an important role in modulating neurite outgrowth and has been demonstrated to be the principal receptor subtype that mediates the protective effects of galanin in the hippocampus (Elliott-Hunt et al. 2007). Millipore's GAL2 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 GAL2 receptor interactions. The membrane preparations exhibit a Kd of 1.4 nM for [125I]-Galanin. With 10 µg/well GAL2 receptor Membrane Prep and 0.5 nM [125I]-Galanin, a greater than 20-fold signal-to-background ratio was obtained.
References
Product Information
Format
Membranes
Presentation
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.
Galanin is an important neuromodulator present in the brain, gastrointestinal system, and hypothalamopituitary axis. It is a 30-amino acid non-C-terminally amidated peptide that potently stimulates growth hormone secretion, inhibits cardiac vagal slowing of heart rate, abolishes sinus arrhythmia, and inhibits postprandial gastrointestinal motility. The actions of galanin are mediated through interaction with specific membrane receptors that are members of the 7-transmembrane family of G protein-coupled receptors. GALR2 interacts with the N-terminal residues of the galanin peptide. The primary signaling mechanism for GALR2 is through the phospholipase C/protein kinase C pathway (via Gq), in contrast to GALR1, which communicates its intracellular signal by inhibition of adenylyl cyclase through Gi. However, it has been demonstrated that GALR2 couples efficiently to both the Gq and Gi proteins to simultaneously activate 2 independent signal transduction pathways.
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.
Incubation 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. 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.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Table 1. Signal:background and specific binding values obtained in a competition binding assay with GAL2 receptor membrane prep and cold galanin (2-29).
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
HTS186M
04053252510588
Documentation
Literatur
Übersicht
Pub Med ID
Activation of the galanin receptor 2 (GalR2) protects the hippocampus from neuronal damage Elliott-Hunt, Caroline R, et al J Neurochem, 100:780-9 (2007)
2007
The neuropeptide galanin, which is widely expressed in brain and peripheral tissues, exerts a broad range of physiological effects. Pharmacological studies using peptide analogues have led to speculation about multiple galanin receptor subtypes. Since 1994, a total of three G-protein-coupled receptor (GPCR) subtypes for galanin have been cloned (GAL1, gal2 and gal3). Potent, selective antagonists are yet to be found for any of the cloned receptors. Major challenges in this field include linking the receptor clones with each of the known physiological actions of galanin and evaluating the evidence for additional galanin receptor subtypes.
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 >>
