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激活STAT蛋白抑制因子3(PIAS3),也被称为E3 SUMO-蛋白连接酶 PIAS3,是由PIAS3基因编码的酶。最初,PIAS3被发现与白细胞介素-6(IL-6)活化的骨髓母细胞M1细胞中的磷酸化STAT3特异性相互作用。这种相互作用是通过PIAS3与STAT3 DNA结合结构域的介导的。因此,STAT3转录活性受到物理预防其与靶基因结合的抑制。随后,它还被发现是其他关键转录因子的调节蛋白,包括MITF,NFκB,SMAD和雌激素受体。它还具有SUMO(小泛素样修饰剂)-E3连接酶的作用,其催化SUMO蛋白与特定靶底物的共价附着。它直接与几种转录因子结合,并阻断或增强它们的活性。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有人激活STAT蛋白抑制因子3(PIAS3)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激活STAT蛋白抑制因子3(PIAS3)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 PIAS3. 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 PIAS3 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.