- 预期应用
- ELISA法定量测定人血清、血浆或其它相关液体中1,25(OH)2D3含量。
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- 实验原理
- 本试剂盒应用双抗体夹心酶标免疫分析法测定标本中1,25(OH)2D3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1,25(OH)2D3、生物素化的抗人1,25(OH)2D3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1,25(OH)2D3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。
- Intended use
- This immunoassay kit allows for the specific measurement of human 1,25-(OH)2D3 concentrations in serum and plasma.
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- Introduction
- 1,25-(OH)2D3, the active metabolite of vitamin D, regulates immune responses in addition to its role in calcium, phosphorus, and bone metabolism. Recent data suggests that 1,25-(OH)2D3 blocks dendritic cell maturation influencing the development of regulatory T cells. We have shown that high dose 1,25-(OH)2D3 monotherapy is effective at delaying acute rejection. We and others have shown that the vitamin D receptor (VDR), which functions as a transcription factor, forms a complex with Smad 3, a TGFb-1 signaling protein, in rat renal lysates for treated recipients. We have now shown that 1,25-(OH)2D3 significantly prolonged graft survival, limited the degree of intersitial fibrosis and glomerulosclerosis, decreased urinary protein and altered Smad and MMP expression in a rat renal model of CAN. We have also observed, in a retrospective clinical study, that cadaveric renal transplant recipients with renal insufficiency placed on calcitriol demonstrate improved renal function and improved graft survival. These results suggest that 1,25-(OH)2D3 may regulate TGFb-1-mediated ECM deposition. Therefore, we hypothesize that 1,25-(OH)2D3 may be an effective agent in preventing or limiting CAN. Further, we hypothesize that 1,25-(OH)2D3 may alter the balance of matrix deposition and removal in mesangial cells by altering MMP expression.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for 1,25-(OH)2D3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any 1,25-(OH)2D3 present is bound by the immobilized antibody. An
enzyme-linked monoclonal antibody specific for 1,25-(OH)2D3 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 1,25-(OH)2D3 bound in the initial step. The color
development is stopped and the intensity of the color is measured. |
