ELISA法定量测定大鼠血清、血浆、细胞培养物上清或其它相关液体中CXCR3含量。

 

本试剂盒应用双抗体夹心酶标免疫分析法测定标本中CXCR3水平。用纯化的抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入CXCR3抗原、生物素化的抗大鼠CXCR3抗体、HRP标记的亲和素，经过彻底洗涤后用底物TMB显色。TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的CXCR3呈正相关。用酶标仪在450nm波长下测定吸光度（OD值），计算样品浓度。

 

This immunoassay kit allows for the specific measurement of rat CXCR3 concentrations in serum and plasma.

 

CD183 (CXCR3) is a multi-pass type-3, 7 span 368 aa glycoprotein.  It contains an extracellular domain which contains 3 potential amino-glycosylation sites, 7 putative transmembrane-spanning domains and an intracellular carboxyl-terminal regions which has 3 intracellular loop regions.  The intracellular carboxyl-terminal region contains 10 Thr/Ser residues that may become phosphorylated during CD183-mediated signaling.  The DRYLAIVHA-motif in the 2nd intracellular loop is the most highly conserved sequence element in chemokine receptors.  CD183 is a chemokine receptor which are G-proteins coupled receptors.  CD183 shares a 30% identity with CD181 (CXCR1) and CD182 (CXCR2).
CD183 has a specificity for 3 chemokines, termed IP10 IFN-g  -inducible 10 kDa protein, termed Mig monokine induced by IFN-r and termed I-TAC interferon-inducible T cell a-chemoattractant.  Nomenclature for chemokines and chemokine receptors is reviewed in structural subfamily CXC chemokines, in which 1 aa residue separates the 1st 2 of 4 highly conserved Cys residues.  Historically, CD183 is the 3rd CXC chemokine receptor discovered and, therefore, commonly designated as CXCR3.  Binding of chemokines to CD183 induces cellular responses that are involved in leukocyte traffic, most notably integrin activation, cytoskeletal changes and chemotactic migration.  Inhibition by Bordetella pertussis toxin suggests the heterotrimeric G protein of the Gi-subclass couple to CD183.  Signal transduction has not been further analyzed but may include the same enzymes including phospholipases, protein kinase B and C, phosphatidylinositol-3-OH kinases, MAP kinases, G protein-coupled receptor kinases, and small GTPases that were identified in the signaling cascade induced by other chemokine receptors.  As a consequence of chemokine-induced cellular desensitization and phosphorylation-dependent receptor internalization, cellular responses are typically rapid and short in duration.  Cellular responsiveness is restored after dephosphorylation of intracellular receptors and subsequent recycling to the cell surface

 

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for CXCR3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CXCR3 present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for CXCR3 is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of CXCR3 bound in the initial step. The color development is stopped and the intensity of the color is measured.