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非组织特异性碱性磷酸酶(ALPL)是一种由ALPL基因编码的酶 。它是一种膜结合的糖基化酶,不在任何特定组织中表达,因此被称为该酶的组织非特异性形式。它与一种被称为低磷酸血症的疾病有直接联系,这种疾病的特点是血清ALP低和骨矿化不足(骨质疏松症)。症状的严重程度取决于ALPL基因中哪个氨基酸发生了变化。ALPL基因的突变导致组织非特异性碱性磷酸酶(TNSALP)的低活性,从而导致低磷酸血症(HPP)。HPP有不同的临床形式,可通过常染色体隐性遗传或常染色体显性遗传。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有小鼠非组织特异性碱性磷酸酶(ALPL)捕获抗体的微孔中,依次加入样本、标准品、生物素标记的检测抗体,HRP酶结合物,中间经过温育和洗涤,用底物TMB显色,TMB在过氧化物酶(HRP)的催化下转化成蓝色,并在酸的作用下转化成最终的黄色。颜色的深浅和样本中的小鼠非组织特异性碱性磷酸酶(ALPL)呈正相关。用酶标仪在450nm 波长下测定吸光度(OD值),计算样本浓度。
This kit employs the sandwich enzyme-linked immunosorbent assay (ELISA) principle. Microplate wells are pre-coated with a capture antibody specific to Mouse ALPL. Following this, the sample, standards, a biotin-conjugated detection antibody, and an HRP (horseradish peroxidase)-labeled conjugate are added sequentially to the wells. The procedure involves incubation and wash steps between each addition. For detection, TMB substrate is used. TMB yields a blue color in the presence of the peroxidase activity of HRP. The reaction is then stopped with an acid solution, which converts the blue color to a final yellow color. The intensity of the yellow color is directly proportional to the concentration of Mouse ALPL present in the sample. The optical density (OD) is measured at a wavelength of 450 nm using a microplate reader. Finally, the concentration of the target analyte in the sample is calculated based on the standard curve generated from the standards.