Key Specifications Table
| Species | Host Cells | Protein Target |
|---|---|---|
| Human | Chem-1 | VPAC2 / VIP2 |
| Description | |
|---|---|
| Catalogue Number | HTS079M |
| Brand Family | Chemicon® |
| Trade Name |
|
| Description | ChemiSCREEN™ Human VPAC2 VIP Receptor Membrane Preparation |
| Overview | Human VPAC2 |
| Background Information | Vasoactive intestinal peptide (VIP), a 28 amino acid peptide originally isolated by its vasodilation activity, binds to two class B GPCRs, VPAC1 and VPAC2, to exert its functions in the CNS, vasculature, immune system and adrenal medulla (Harmar et al., 1998). In the immune system, VPAC2 is expressed on stimulated CD4 T cells, and binding of T cell-derived VIP to VPAC2 induces a shift toward the Th2 pathway (Delgado et al., 2004; Voice et al., 2004). In addition, VPAC2 is an essential regulator of the circadian pacemaker of the hypothalamic suprachiasmatic nuclei (Hughes et al., 2004). Chemicons VPAC2 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 VPAC2 interactions with PACAP. The membrane preparations exhibit a Kd of 0.9 nM for [125I]-PACAP27. With 5 µg/well VPAC2 Membrane Prep and 0.5 nM [125I]-PACAP27, a greater than 15-fold signal-to-background ratio was obtained. |
| Product Information | |
|---|---|
| Format | Membranes |
| Presentation | 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. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Human VPAC2 / VIP2 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 | Vasoactive intestinal peptide (VIP; MIM 192320) and pituitary adenylate cyclase activating polypeptide (PACAP; MIM 102980) are homologous peptides that function as neurotransmitters and neuroendocrine hormones. While the receptors for VIP and PACAP share homology, they differ in their substrate specificities and expression patterns. See VIPR1 (MIM 192321) and ADCYAP1R1(MIM 102981).[supplied by OMIM] |
| Gene Symbol |
|
| GPCR Class | B |
| Host Cells | Chem-1 |
| Protein Target | VPAC2 / VIP2 |
| Target Sub-Family | VIP/PACAP |
| UniProt Number |
|
| UniProt Summary | FUNCTION: SwissProt: P41587 # This is a receptor for VIP as well as PACAP-38 and -27, the activity of this receptor is mediated by G proteins which activate adenylyl cyclase. Can be coupled to phospholipase C. SIZE: 438 amino acids; 49479 Da SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein. SIMILARITY: SwissProt: P41587 ## Belongs to the G-protein coupled receptor 2 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, a GF/C 96-well filter plate 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] PACAP27 (Perkin Elmer # NEX294) Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C. One package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 15-fold signal:background with 125I-labeled PACAP27 at 0.5 nM. |
| Product Usage Statements |
|---|
| 10 µg/well | 5 µg/well | |
|---|---|---|
| Signal:Background | 25.5. | 22.8 |
| Specific Binding (cpm) | 32913 | 27650 |
SPECIFICATIONS: 1 unit = 5 µg membrane preparation
Bmax: 17.3 pmol/mg
Kd: 0.9 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 |