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48-602MAG
Buffer Detection Kit for Magnetic Beads
1 Kit
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HTS074M
Sigma-AldrichChemiSCREEN™ Human PK1 Prokineticin Receptor Membrane Preparation
Human PK1 / PKR1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
ChemiSCREEN™ Human PK1 Prokineticin Receptor Membrane Preparation
Overview
Full-length human GPR73 cDNA, encoding PK1
Background Information
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 Gq-coupled receptors, PK1 and PK2 (also known as GPR73a and GPR73b), mediate cellular responses to prokineticins (Lin et al., 2002). Chemicon's PK1 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 PK1 interactions and its ligands. The membrane preparations exhibit a Kd of 0.11 nM for [125I]-Mamba Intestinal Toxin-1 (MIT-1). With 0.2 nM [125I]-MIT-1, 2.5 µg/well and 5 µg/well PK1 Membrane Prep typically yield greater than 5-fold signal-to-background ratio.
References
Product Information
Format
Membranes
Presentation
Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives. Packaging method: Membranes protein were adjusted to 0.5 mg/ml in 1 mL packaging buffer, rapidly frozen, and stored at -80°C.
FUNCTION: SwissProt: Q8TCW9 # Receptor for prokineticin 1. Exclusively coupled to the G(q) subclass of heteromeric G proteins. Activation leads to mobilization of calcium, stimulation of phosphoinositide turnover and activation of p44/p42 mitogen-activated protein kinase. SIZE: 393 amino acids; 44770 Da SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein. TISSUE SPECIFICITY: Expressed in the stomach, throughout the small intestine, colon, rectum, thyroid gland, pituitary gland, salivary gland, adrenal gland, testis, ovary, brain, spleen, prostate and pancreas. SIMILARITY: SwissProt: Q8TCW9 ## 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, 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]-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. One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 10-fold signal:background with 125I-labeled MIT-1 at 0.2 nM
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of PK1 membrane prep:
5 µg/well
2.5 µg/well
Signal:Background
6
6.4
Specific Binding (cpm)
3166
1798
SPECIFICATIONS: 1 unit = 5 µg Bmax for [125I] MIT-1 binding: 0.58 pmol/mg protein; Kd for [125I] MIT-1 binding: ~ 0.11 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
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Bestellnummer
GTIN
HTS074M
04053252326592
Documentation
ChemiSCREEN™ Human PK1 Prokineticin Receptor Membrane Preparation SDB
The suprachiasmatic nucleus (SCN) controls the circadian rhythm of physiological and behavioural processes in mammals. Here we show that prokineticin 2 (PK2), a cysteine-rich secreted protein, functions as an output molecule from the SCN circadian clock. PK2 messenger RNA is rhythmically expressed in the SCN, and the phase of PK2 rhythm is responsive to light entrainment. Molecular and genetic studies have revealed that PK2 is a gene that is controlled by a circadian clock (clock-controlled). Receptor for PK2 (PKR2) is abundantly expressed in major target nuclei of the SCN output pathway. Inhibition of nocturnal locomotor activity in rats by intracerebroventricular delivery of recombinant PK2 during subjective night, when the endogenous PK2 mRNA level is low, further supports the hypothesis that PK2 is an output molecule that transmits behavioural circadian rhythm. The high expression of PKR2 mRNA within the SCN and the positive feedback of PK2 on its own transcription through activation of PKR2 suggest that PK2 may also function locally within the SCN to synchronize output.
Endocrine gland-derived VEGF and the emerging hypothesis of organ-specific regulation of angiogenesis. LeCouter, Jennifer, et al. Nat. Med., 8: 913-7 (2002)
2002
The diversity in growth and morphological characteristics among endothelial cells in different normal tissues and tumors has been long recognized. Yet there has been no clear molecular explanation for such diversity at the level of vascular endothelial growth factor A (VEGF-A) and other established regulators of angiogenesis that are expressed widely and show little tissue selectivity in their angiogenic properties. Endocrine gland-derived VEGF represents the first example of a tissue-specific angiogenic factor, likely to be followed by others.
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 >>
