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泛素特异性肽酶14(USP14),也被称为泛素硫酯酶14或去泛素化酶14,是由USP14基因编码的酶。该基因编码泛素特异性加工(UBP)蛋白酶家族的成员,该蛋白酶是一种具有His和Cys结构域的去泛素化酶(DUB)。它位于细胞质中,并切割从泛素前体融合和泛素化蛋白质泛素部分。它是趋化因子受体CXCR4降解所必需的,这对于CXCL12诱导的细胞趋化性至关重要。它通过与ERN1相互作用抑制未折叠内质网蛋白的降解,还可作为非应激条件下内质网相关降解(ERAD)的生理抑制剂。它对于神经肌肉接头(NMJ)的突触发育和功能必不可少。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有人泛素特异性肽酶14(USP14)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人泛素特异性肽酶14(USP14)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 Human USP14. 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 Human USP14 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.