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硫酸乙酰肝素酶(HPSE),是一种在细胞表面和细胞外基质内发挥作用的酶,可将聚合硫酸肝素分子降解为较短链长的寡糖。该蛋白最初在高尔基体中以非活性的 65kDa 原肝素酶的形式合成,并转移到晚期内体/溶酶体中以运输到细胞表面。在溶酶体中,它被蛋白酶加工成其活性形式。它具有内切糖苷酶的活性,在聚合链内部的部位裂解聚合的硫酸肝素分子。该酶降解基底膜和细胞外基质的硫酸肝素支架。它还与炎症过程有关,允许活化的 T 淋巴细胞外渗。在眼球表面的生理学中,这种活性是前分泌性有丝分裂蛋白的一个开关。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有小鼠硫酸乙酰肝素酶(HPSE)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硫酸乙酰肝素酶(HPSE)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 HPSE. 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 HPSE 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.