Key Specifications Table
| Species | Host Cells | Protein Target |
|---|---|---|
| Human | Chem-2 | H2 |
| Description | |
|---|---|
| Catalogue Number | HTS086M |
| Brand Family | Chemicon® |
| Trade Name |
|
| Description | ChemiSCREEN™ Human H2 Histamine Receptor Membrane Preparation |
| Overview | Full-length human HRH2 cDNA encoding H2 |
| Background Information | The histamine H2 receptor is a widely distributed G-protein coupled receptor, which is important in regulating a host of physiologic actions extending from gastric acid secretion to gastrointestinal motility (Del Valle and Gantz, 1997). Histamine mediated activation of its receptor leads to signaling through both adenylate cyclase and phosphoinositide /protein kinase C second messenger systems (Hill, 1990). Antagonists for this receptor have proven to be effective therapy for acid peptic disorders of the GI tract. Certain antagonists are used in the treatment of neuropsychiatric and neurological diseases e.g. schizophrenia, Alzheimers disease and Parkinsons disease. Chemicons H2 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 at H2. The membrane preparations exhibit a Kd of 0.8 nM for [125I]-Iodoaminopotentidine. With 0.65 nM [125I]-Iodoaminopotentidine, 10 µg/well H2 Membrane Prep typically yields greater than 7-fold signal-to-background ratio. |
| Product Information | |
|---|---|
| Format | Membranes |
| Presentation | One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 7-fold signal:background with 125I-labeled Iodoaminopotentidine at 0.65 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 1 mg/mL in packaging buffer, and dispensed at 1 mL/vial. Vials were rapidly frozen, and stored at -80oC. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Human H2 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding. |
| Key Applications |
|
| Application Notes | Radioligand binding assay and GTPγS binding. |
| Biological Information | |
|---|---|
| Entrez Gene Number |
|
| Entrez Gene Summary | Histamine is a ubiquitous messenger molecule released from mast cells, enterochromaffin-like cells, and neurons. Its various actions are mediated by histamine receptors H1, H2, H3 and H4. Histamine receptor H2 belongs to the family 1 of G protein-coupled receptors. It is an integral membrane protein and stimulates gastric acid secretion. It also regulates gastrointestinal motility and intestinal secretion and is thought to be involved in regulating cell growth and differentiation. |
| Gene Symbol |
|
| GPCR Class | A |
| Host Cells | Chem-2 |
| Protein Target | H2 |
| Target Sub-Family | Histamine |
| UniProt Number |
|
| UniProt Summary | FUNCTION: SwissProt: P25021 # The H2 subclass of histamine receptors mediates gastric acid secretion. Also appears to regulate gastrointestinal motility and intestinal secretion. Possible role in regulating cell growth and differentiation. The activity of this receptor is mediated by G proteins which activate adenylyl cyclase and, through a separate G protein-dependent mechanism, the phosphoinositide/protein kinase (PKC) signaling pathway (By similarity). SIZE: 359 amino acids; 40098 Da SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein. SIMILARITY: SwissProt: P25021 ## Belongs to the G-protein coupled receptor 1 family. MISCELLANEOUS: Antagonists for this receptor have proven to be effective therapy for acid peptic disorders of the gastrointestinal tract. Certain antagonists are used in the treatment of neuropsychiatric and neurological diseases such as schizophrenia, Alzheimer disease and Parkinson disease. |
| 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, 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] Iodoaminopotentidine (Amersham#: IM264) Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C. |
| Product Usage Statements |
|---|
| 10 µg/well | |
|---|---|
| Signal:Background | 11.68 |
| Specific Binding (cpm) | 4152 |
SPECIFICATIONS: 1 unit = 10 µg
Bmax for [125I] Iodoaminopotentidine binding: 0.46 pmol/mg protein
Kd for [125I] Iodoaminopotentidine binding: ~0.8 nM 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.
| 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 |