HTS182M

ChemiSCREEN Membrane Preparation Recombinant Human PK ₂ Prokineticin Receptor

• UNSPSC-Code: 41106514
• eCl@ss: 32161000
• NACRES: NA.45

Eigenschaften

• Biologische Quelle: human
• Qualitätsniveau: 100
• Rekombinant: expressed in Chem-1 cells
• Form: liquid
• Hersteller/Markenname: ChemiScreen; Chemicon^®
• Methode(n): ligand binding assay: suitable (GTPγS); radioligand binding assay (RLBA): suitable
• NCBI-Hinterlegungsnummer: NM_144773.2
• UniProt-Hinterlegungsnummer: Q8NFJ6

Beschreibung

Allgemeine Beschreibung

Full-length human GPR73L1 cDNA encoding PK₂

Prokineticins, also known as endocrine gland vascular endothelial growth factors (EG-VEGF), are two ~10 kD secreted proteins originally described to mediate angiogenesis and gastrointestinal smooth muscle contraction (Li et al., 2001; LeCouter et al., 2003). Subsequently, prokineticins have been found to mediate central nervous system functions including circadian rhythms and olfactory bulb development (Cheng et al., 2002; Ng et al., 2005). Two G_q-coupled receptors, PK₁ and PK₂ (also known as GPR73a and GPR73b), mediate cellular responses to prokineticins (Lin et al., 2002). Millipore′s PK₂ 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 PK₂. The membrane preparations exhibit a K_d of 0.36 nM for [¹²⁵I]-MIT-1. With 0.3 nM [¹²⁵I]-MIT-1, 10μg/well PK₂ Membrane Prep typically yields greater than 5-fold signal-to-background ratio.

Anwendung

Human PK2 / PKR2 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

Radioligand binding assay and GTPγS binding.

Biochem./physiol. Wirkung

• GPCR Class: A
• Protein Target: PK2/PKR2
• Target Sub-Family: Prokineticin

Spezifikationen

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] MIT-1 (Perkin Elmer#: NEX-410)

Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.

Physikalische Form

One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 5-fold signal:background with ¹²⁵I-labeled MIT-1 at 0.3 nM

Liquid in packaging buffer: 50 m

Lagerung und Haltbarkeit

Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.

Rechtliche Hinweise

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Haftungsausschluss

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

Sicherheitshinweise

• Lagerklassenschlüssel: 12 - Non Combustible Liquids
• WGK: WGK 2