HTS109C | ChemiScreen™ Human 5-HT2B Serotonin Receptor Calcium-Optimized FLIPR Cell Line

Calcium flux in 5-HT2B-expressing CHO cell line induced by α-methyl- 5-hydroxytryptamine. 5-HT2B-expressing CHO cells and Wild-Type CHO cells were loaded with Fluo-4 NW and calcium flux in response to α-methyl- 5-hydroxytryptamine (10-7 to 10-12.5 M) was determined in triplicate on a Molecular Devices FLIPRTETRA™.
Calcium flux in 5-HT2B-expressing CHO cell line induced by α-methyl- 5-hydroxytryptamine.
HTS109C
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      Description
      Catalogue Number HTS109C
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      • ChemiScreen
      Description ChemiScreen™ Human 5-HT2B Serotonin Receptor Calcium-Optimized FLIPR Cell Line
      Overview Full-length human HTR2B cDNA encoding 5-HT2B
      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). The 5-HT2B receptor, which couples to Gq/11 to increase intracellular calcium, is expressed in embryonic and adult cardiovascular tissues, gut and brain from the rat, mouse, and human species. A role for 5-HT2B receptors was suggested in the cardiopathy associated with fenfluramine (Fitzgerald et al., 2000). Chemicon's cloned human 5-HT2B -expressing cell line is made in the CHO host, which supports high levels of recombinant 5-HT2B expression on the cell surface. Thus, the cell line is an ideal tool for screening for antagonists of interactions between 5-HT2B and its ligands.
      References
      Product Information
      Presentation Cells are frozen at 2 x 106 cells/mL in F-12K Nutrient Mixture, Kaighn's Modification /20% dialyzed heat inactivated fetal bovine serum /10% DMSO. Cell line tests negative for mycoplasma.
      Applications
      Application Human 5-HT2B GPCR stably expressed in CHO cells for Calcium flux FLIPR Assays.
      Application Notes Calcium flux assay, ligand binding assays
      Biological Information
      Species Human
      Species Reactivity Human
      Entrez Gene Number
      Entrez Gene Summary Multiple receptor subtypes of serotonin neurotransmitters with multiple physiologic functions have been recognized. The 5-HT-2 receptors mediate many of the central and peripheral physiologic functions of serotonin. Cardiovascular effects include contraction of blood vessels and shape changes in platelets; central nervous system effects include neuronal sensitization to tactile stimuli and mediation of hallucinogenic effects of phenylisopropylamine hallucinogens.[supplied by OMIM]
      Gene Symbol
      • 5-HT(2B)
      • HTR2B
      • 5-HT-2B
      • 5-HT2B
      GPCR Class A
      Host Cells CHO (Chinese hamster ovary cells)
      Protein Target 5-HT2B
      Target Sub-Family Serotonin
      UniProt Number
      UniProt Summary FUNCTION: SwissProt: P41595 # This is one of the several different receptors for 5- hydroxytryptamine (serotonin), a biogenic hormone that functions as a neurotransmitter, a hormone, and a mitogen. This receptor mediates its action by association with G proteins that activate a phosphatidylinositol-calcium second messenger system.
      SIZE: 481 amino acids; 54298 Da
      SUBUNIT: Interacts with MPDZ.
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Detected in most peripheral organs. Only low expression levels were found in the brain.
      SIMILARITY: SwissProt: P41595 ## Belongs to the G-protein coupled receptor 1 family.
      Cell Line Type
      • GPCR Cell Lines
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality Assurance EC50 for calcium mobilization by α-methyl-5-hydroxytryptamine: ~ 0.35nM
      Signal:noise at agonist Emax: 244
      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.
      • This product contains genetically modified organisms (GMO). Within the EU GMOs are regulated by Directives 2001/18/EC and 2009/41/EC of the European Parliament and of the Council and their national implementation in the member States respectively. This legislation obliges Merck-Millipore to request certain information about you and the establishment where the GMOs are being handled. Click here for Enduser Declaration (EUD) Form.
      Storage and Shipping Information
      Storage Conditions Place cells in liquid nitrogen immediately upon receipt. Maintain frozen in liquid nitrogen for up to 5 years. Thaw cells rapidly by removing from liquid nitrogen and immediately immersing in a 37ºC water bath. Immediately after ice has thawed, sterilize the exterior of the vial with 70% ethanol. Transfer contents of the vial to a T75 flask containing 20 mL growth media, and place in a humidified 37°C incubator with 5% CO2.. After 8-24 h, cells will adhere to the plate, at which time the media should be replaced to remove residual DMSO. Cells are passaged by washing with Ca++ and Mg++-free HBSS (10 mL/T75), incubating with 0.05% trypsin/0.2 g/L EDTA (1 mL/T75) for 5-10 minutes at 37°C, and rapping the side of the flask to dislodge the cells. Neutralize the trypsin by addition of 4 volumes growth media. Cells are typically passaged 1:10 with every 3-4 days, and should be passaged at least once after thawing prior to use in calcium flux assays.
      Packaging Information
      Packaging 1 mL per vial, 2 x 106 cells/mL
      Qty/Pck 2 vials
      Material Size 2
      Material Package vials
      Transport Information
      Supplemental Information
      Specifications

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