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胱天蛋白酶3(Casp-3,Caspase-3)是一种与caspase-8和caspase-9相互作用的caspase蛋白。它是由CASP3基因编码的。CASP3的同源物已在许多有完整基因组数据的哺乳动物中被发现。CASP3蛋白是半胱氨酸-天冬氨酸蛋白酶(caspase)家族的成员。caspases的顺序激活在细胞凋亡的执行阶段起着核心作用。它以非活性原酶的形式存在,经过保守的天冬氨酸残基的蛋白水解处理,产生两个亚单位,大的和小的,二聚体形成活性酶。该蛋白裂解并激活了Caspases 6和7;而该蛋白本身被Caspases 8、9和10加工并激活。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有小鼠胱天蛋白酶3(CASP3)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胱天蛋白酶3(CASP3)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 CASP3. 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 CASP3 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.