ElaBoXTM小鼠核苷酸结合寡聚化结构域样受体蛋白3检测试

试剂盒类型

传统ELISA试剂盒

检测原理

Solarbio ELISA试剂盒采用基于双抗体夹心法的酶联免疫吸附检测技术。将抗小鼠NLRP3单克隆抗体包被在酶标板上；分别加入梯度稀释的标准品和预稀释的样本，标准品和样本中的小鼠NLRP3会与酶标板上的包被抗体充分结合；洗板后加入生物素化抗小鼠NLRP3抗体，该抗体会与板子上包被抗体捕获的标准品和样本中的小鼠NLRP3发生特异性结合；洗板后加入辣根过氧化物酶（HRP）标记的链霉亲和素，生物素与链霉亲和素会发生高强度的非共价结合；洗板后加入显色剂底物TMB，若反应孔中样品存在不同浓度的小鼠NLRP3，则HRP会使无色TMB变成不同深浅（正相关）的蓝色物质，加入终止液后反应孔会变成黄色；最后，在λmax=450 nm（OD=450 nm）处测定反应孔样品吸光度（OD），样本中的小鼠NLRP3浓度与OD成正比，通过绘制标准曲线和四参数拟合软件便可计算出样本中小鼠NLRP3的浓度。

背景说明

NLRP3, also known as NALP3, CIAS1, PYPAF or Cryopyrin, is a cytosolic ~120 kDa member of the NLRP family of proteins expressed in leukocytes, especially neutrophils. As a component of the inflammasome, NLRP3 activates caspases 1 and 5. Defects in NLRP3 may cause FCAS1, CINCA, or Muckle-Wells autoinflammatory syndromes. Mouse NLRP3 contains an N-terminal Pyrin domain (aa 1-91) followed by a Nacht region (aa 216-532), and seven LRRs (aa 739-988). Within the sequence used as an immunogen, mouse NLRP3 shares 78% and 93% aa identity with human and rat NLRP3, respectively. Alternate splicing of mouse NLRP3 generates additional isoforms that lack either LRR2 and 3, LRR 6 and 7, or LRR4-9.