Key Specifications Table
| Species | Host Cells | Protein Target |
|---|---|---|
| Human | Chem-1 | CCR10 |
| Description | |
|---|---|
| Catalogue Number | HTS014M |
| Brand Family | Chemicon® |
| Trade Name |
|
| Description | ChemiSCREEN™ CCR10 Membrane Preparation |
| Overview | Full length human CCR10 with a proprietary mutation in the cytoplasmic domain to enhance cell surface expression. |
| Background Information | CCR10, previously known as GPR2, is a GPCR that is expressed in dermal endothelial cells, fibroblasts, melanocytes, and circulating T cells and PBMCs (Homey et al., 2000). Two chemokines, CTACK/CCL27 and MEC/CCL28, bind to CCR10 and stimulate migration and calcium flux (Pan et al., 2000; Wang et al., 2000). Expression of CTACK by keratinocytes, and CCR10 by lymphocytes from skin of patients with psoriasis and dermatitis, implicates this receptor ligand pair in skin inflammation. In addition, disruption of CCL27-CCR10 interaction reduces skin inflammation induced by allergen (Homey et al., 2002). CCR10 expressed on IgA antibody-secreting cells also mediates their migration to CCL28 expressed in the lactating mammary gland, and subsequent IgA accumulation in milk (Wilson and Butcher, 2004). Millipores CCR10 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 CCR10 interactions with its ligands. The membrane preparations exhibit an EC50 of 47 nM for CTACK/CCL27 in a GTPγS binding assay. |
| 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 -80oC. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Human CCR10 GPCR membrane preparation for GTPγS Binding Assays. |
| Key Applications |
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| Application Notes | GTPγS Binding Assay. |
| Biological Information | |
|---|---|
| Concentration | 1.0 mg/mL |
| Entrez Gene Number |
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| Entrez Gene Summary | The 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 |
|
| GPCR Class | A |
| Host Cells | Chem-1 |
| Protein Target | CCR10 |
| Target Sub-Family | Chemokine |
| UniProt Number |
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| UniProt Summary | FUNCTION: 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 Conditions | GTPγS ASSAY CONDITIONS: Membranes are permeabilized by addition of saponin to an equal concentration by mass, then mixed with [35S]-GTPγS (final concentration of 0.3 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 ligand was 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 an FB filter plate (Millipore MAHF B1H) previously prewetted with water. The plate is washed 3 times (1 mL per well per wash) with cold 10 mM sodium phosphate, pH 7.4, then dried and counted. |
| Product Usage Statements | |
|---|---|
| Quality Assurance | EC50 for CTACK/CCL27: ~5.6 nM in a GTPγS binding assay |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Store at –70°C. Product is stable for at least 6 months from the date of receipt when stored as directed. Do not freeze and thaw. |
| Packaging Information | |
|---|---|
| Material Size | 200 assays |