【项目简介】
Cytochrome c was identified as a component required for the crucial steps in apoptosis，caspase-3 activation and DNA fragmentation。Cytochrome c was shown to redistribute from mitochondria to cytosol during apoptosis in intact cells。Mitchrondrial cytochrome c is a water-soluble protein of 15 kDa with a net positive charge，residing loosely attached in the mitochrondrial intermembrane space。Cytochrome c functions in the respiratory chain by interaction with redox partners。Measurement of cytochrome c release from the mitochondria is a tool to detect the first early steps for initiating apoptosis in cells。Moreover，elevated cytochrome c levels were observed in serum from patients with hematological malignancies。In the course of cancer chemotherapy，the serum-cytochrome c level grew rapidly and it decreased gradually as the patient was cleared from malignant cells。Thus, serum-cytochrome c monitoring might serve as a clinical marker indicating the onset of apoptosis and cell turn-over in vivo。

【检验原理】
本试剂盒采用双抗体夹心ELISA法。酶标微孔板上预包被有抗人Cytochrome c抗体，当加入标准品和待测样本时，重组或天然的人Cytochrome c会和该抗体结合。洗板后，游离的其他成分被洗去，加入生物素化的抗人Cytochrome c抗体可与被酶标板捕获的人Cytochrome c结合。再次洗板后，游离的其他成分被洗去，加入辣根过氧化物酶标记的链霉亲和素，生物素和链霉亲和素特异性结合，形成“包被抗人Cytochrome c抗体+人Cytochrome c+生物素化抗人Cytochrome c抗体+链霉亲和素-HRP”免疫复合物。当加入TMB显色底物后，复合物上连接的HRP催化底物反应，生成蓝色产物，终止反应后变为黄色。在450nm处测定OD值，人Cytochrome c浓度与OD450值之间呈正比，可通过绘制标准曲线求出标本中人Cytochrome c的浓度。