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HTS036M ChemiSCREEN™ CCR9 Membrane Preparation

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HTS036M
1 mL  
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      Overview

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1CCR9
      Description
      Catalogue NumberHTS036M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ CCR9 Membrane Preparation
      OverviewFull length Human CCR9
      Background InformationThe CCR9 receptor is a GPCR expressed selectively in the thymus, lymph node, spleen, and the small bowel (SB). CCR9 binds to Thymus-expressed chemokine ligand (TECK / CCL25) (Zaballos et al. 1999; Youn et al., 1999; Yu et al., 2000), and is involved in directing the movement of mature T cells in the thymus (Carramolino et al., 2001b). CCR9 also plays an important role in the pathogenesis of SB Crohn's disease (Carramolino et al., 2001a; Papadakis et al, 2001). The CCR9 selective expression in the small bowel but not the colon indicates the role of CCR9 in the specialized immune response in the SB (Kunkel et al., 2000; Papadakis et al., 2000). It was also shown that CCR9-mediated antiapoptosis could help in the study of cell survival mechanism and T cell malfunctioning (Youn et al, 2002). Chemicon's CCR9 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 CCR9 interactions with its ligands. The membrane preparations exhibit an EC50 of 8 nM for TECK/CCL25 in a GTPgammaS binding assay.
      References
      Product Information
      FormatMembranes
      PresentationLiquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.Packaging method: Membrane protein was adjusted to 0.5 mg/ml in packaging buffer, rapidly frozen, and stored at -80°C.
      Quality LevelMQ100
      Applications
      ApplicationHuman CCR9 GPCR membrane preparation for GTPγS Binding & Radioligand Binding Assays.
      Key Applications
      • GTPγS binding
      • Radioligand binding assay
      Application NotesGTPγS Binding and Radioligand Binding Assay.
      Biological Information
      Concentration0.5 mg/mL
      Entrez Gene Number
      Entrez Gene SummaryThe protein encoded by this gene is a member of the beta chemokine receptor family. It is predicted to be a seven transmembrane protein similar to G protein-coupled receptors. Chemokines and their receptors are key regulators of the thymocytes migration and maturation in normal and inflammation conditions. The specific ligand of this receptor is CCL25. It has been found that this gene is differentially expressed by T lymphocytes of small intestine and colon, suggested a role in the thymocytes recruitment and development that may permit functional specialization of immune responses in different segment of the gastrointestinal tract. This gene is mapped to the chemokine receptor gene cluster region. Two alternatively spliced transcript variants have been described.
      Gene Symbol
      • CCR9
      • GPR28
      • CC-CKR-9
      • D6
      • GPR-9-6
      • CDw199
      • CCR10
      • CCR-9
      • CMKBR9
      • CCBP2
      GPCR ClassA
      Host CellsChem-1
      Protein TargetCCR9
      Target Sub-FamilyChemokine
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P51686 # Receptor for chemokine SCYA25/TECK. Subsequently transduces a signal by increasing the intracellular calcium ions level. Alternative coreceptor with CD4 for HIV-1 infection.
      SIZE: 369 amino acids; 42016 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Highly expressed in the thymus and low in lymph nodes and spleen.
      SIMILARITY: SwissProt: P51686 ## Belongs to the G-protein coupled receptor 1 family.
      MISCELLANEOUS: EC50 of SCYA25/TECK for isoform 1 is lower than for isoform 2.
      Incubation ConditionsASSAY CONDITIONS: Membranes are permeabilized by addition of saponin to an equal concentration by mass, then mixed with [35S]-GTPγS (final concentration of 0.1 nM) in 20 mM HEPES, pH 7.4/100 mM NaCl/10 mM MgCl2/0.5 μM GDP in a nonbinding 96-well plate. Unlabeled TECK/CCL25 is added to the final concentration indicated in Figure 1 (final volume 100 μL), and incubated for 30 min at 30°C. The binding reaction is transferred to a GF/B filter plate (prewetted with water), and washed 3 times (1 mL per well per wash) with cold 10 mM sodium phosphate, pH 7.4. The plate is dried and counted.

      One vial contains enough membranes for at least 200 assays (units), where one unit is the amount of membrane that will yield greater than 1000 cpm specific TECK-stimulated [35S]-GTPγS binding.

      The CCR9 membrane preparation is expected to be functional in a radioligand binding assay; however, the end user will need to determine the optimal radiolabeled ligand for use with this product.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceSPECIFICATIONS:
      Kd for TECK/CCL25: ~8 nM in a GTPγS binding assay
      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 ConditionsMaintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
      Packaging Information
      Material Size1 mL
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      HTS036M 04053252677274

      Documentation

      ChemiSCREEN™ CCR9 Membrane Preparation MSDS

      Title

      Safety Data Sheet (SDS) 

      ChemiSCREEN™ CCR9 Membrane Preparation Certificates of Analysis

      TitleLot Number
      CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN CCR9 CHEMOKINE RECEPTOR - 2089073 2089073

      References

      Reference overviewPub Med ID
      Role of the CC chemokine receptor 9/TECK interaction in apoptosis.
      Youn BS, et al.
      Apoptosis, 7(3):271-6 (2002)  2002

      11997671 11997671
      Expression of CCR9 beta-chemokine receptor is modulated in thymocyte differentiation and is selectively maintained in CD8(+) T cells from secondary lymphoid organs.
      Carramolino L, et al.
      Blood, 15;97(4):850-7 (2001)  2001

      11159507 11159507
      The role of thymus-expressed chemokine and its receptor CCR9 on lymphocytes in the regional specialization of the mucosal immune system.
      Papadakis KA, et al.
      J. Immunol., 165(9):5069-76 (2000)  2000

      11046037 11046037
      Lymphocyte CC chemokine receptor 9 and epithelial thymus-expressed chemokine (TECK) expression distinguish the small intestinal immune compartment: Epithelial expression of tissue-specific chemokines as an organizing principle in regional immunity.
      Kunkel, E J, et al.
      J. Exp. Med., 192: 761-8 (2000)  2000

      Show Abstract
      10974041 10974041
      TECK, an efficacious chemoattractant for human thymocytes, uses GPR-9-6/CCR9 as a specific receptor.
      Youn BS, et al.
      Blood, 94(7):2533-6 (1999)  1999

      10498628 10498628

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      Categories

      Life Science Research > Drug Discovery and Development > GPCR Cell Lines & Assays > GPCR Membrane Preparations
      Life Science Research > Proteins and Enzymes > GPCR Membrane Preparations