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Competition binding for 5-HT4B. 5-HT4B Membrane Preparation (5 and 10 μg/well) and 10 μg/well Wild-Type Chem-1 membrane prepara...
Rank ordering of compounds in competition binding for 5-HT4B. 5-HT4B Membrane Preparation (5 μg/well) was incubated with 1 nM [...
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ChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT4B Serotonin Receptor
Description:
ChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT4B Serotonin Receptor
Trade Name:
Chemicon (Millipore)
Qty/Pk:
200 units
Product Overview:
Human HTR4, transcript b, encoding 5-HT4B
Background Information:
5-Hydroxytryptamine (5-HT, also commonly known as serotonin) is synthesized in enterochromaffin cells in the intestine and in serotonergic nerve terminals. In the periphery, 5-HT mediates gastrointestinal motility, platelet aggregation, and contraction of blood vessels. Many functions of the central nervous system are influenced by 5-HT, including sleep, motor activity, sensory perception, arousal and appetite. A family of 12 GPCRs and one ion channel mediate the biological effects of 5-HT (Hoyer et al., 1994). 5-HT4 comprises at least 8 isoforms varying at the C-terminus, which are generated by alternative splicing. The expression and distribution of these splice variants differs among organs and tissues with many of them present in several tissues such as atrium, brain, and GI tract (Bockaert et al. 2004). To date, all isoforms have been shown to activate adenylyl cyclase in vitro, and no difference in signal transduction between C-terminal 5-HT4 receptor variants has been demonstrated. 5-HT4 receptors are of potential interest for the treatment of patients with GI motility disorders and Alzheimer's disease (De Maeyer et al. 2008; Lezoualc’h 2007). Millipore's 5-HT4B 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 5-HT4B. The membrane preparations exhibit a Kd of 0.46 nM for [3H]-GR113808. With 1 nM [3H]-GR113808, 5 μg/well 5-HT4B Membrane Prep yields greater than 10 fold signal-to-background ratio.
Application Notes:
Radioligand binding assay and GTPγS binding.
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.
Species:
Human
Quality Assurance:
Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of 5-HT4B membrane prep.
SPECIFICATIONS: 1 unit = 5 μg membrane preparation
Bmax: 12.5 pmol/mg
Kd: 2.7 nM for [3H]-GR113808
| 10 µg/well | 5 µg/well | |
|---|---|---|
| Signal:Background | 26.4 | 21.7 |
| Specific Binding (cpm) | 5286 | 4879 |
SPECIFICATIONS: 1 unit = 5 μg membrane preparation
Bmax: 12.5 pmol/mg
Kd: 2.7 nM for [3H]-GR113808
Cell Type:
Chem-1
Brand Family:
Chemicon
Incubation Conditions:
RECOMMENDED ASSAY CONDITIONS: Membranes are mixed with radioactive ligand and unlabeled competitor (see Figures 1 and 2 for concentrations tested) in assay 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 assay buffer. Binding reaction is transferred to the filter plate, and washed 3 times (1 mL per well per wash) with assay buffer. The plate is dried and counted.
Assay buffer: 25 mM Tris, pH 7.4, filtered and stored at 4°C
Radioligand: [3H]-GR113808 (Amersham TRK944)
One package contains enough membranes for at least 200 assays (units), where one unit is the amount of membrane that will yield greater than 10-fold signal:background with 3H-labeled GR113808 at 1 nM.
Assay buffer: 25 mM Tris, pH 7.4, filtered and stored at 4°C
Radioligand: [3H]-GR113808 (Amersham TRK944)
One package contains enough membranes for at least 200 assays (units), where one unit is the amount of membrane that will yield greater than 10-fold signal:background with 3H-labeled GR113808 at 1 nM.
Host Cells:
Chem-1
Presentation:
Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
Packaging method: Membrane protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
Packaging method: Membrane protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
Protein Target:
5-HT4
Format:
Membranes
Ligand Type:
non-peptide
Storage Conditions:
Store at –70°C. Product is stable for at least 6 months from the date of receipt when stored as directed. Do not freeze and thaw.
Target Sub-Family:
Serotonin
Promotional Text:
For special offers Click here http://www.millipore.com/drugdiscovery/dd3/gpcrtargetsolutions
GPCR Class:
A
Protein or Enzyme Type:
GPCR Membrane Preparations
Product Name:
ChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT4B Serotonin Receptor
Entrez Gene Number:
Cell Membrane Type:
GPCR