辅酶Ⅰ NAD（H）含量检测试剂盒

检测样本量： 25 Samples
主要检测设备及配套：可见分光光度计/1 mL玻璃比色皿（d=10 mm）
预计测定时间： 4 h (25 Samples)
需自备试剂： 无水乙醇（C₂H₆O，MW=46.07，CAS: 64-17-5）
试剂储存条件： 按照标签要求储存

通过酸性和碱性提取液分别提取样品中NAD⁺和NADH，NADH可通过PMS的递氢作用，还原氧化型噻唑蓝（MTT）生成甲瓒，产物在570 nm处具有特征吸收峰；NAD⁺可被乙醇脱氢酶还原为NADH，进一步采用MTT还原法生成甲瓒，通过吸光值变化即可定量检测辅酶Ⅰ NAD (H) 的含量。

• 检测方法： MTT还原法
• 检测波长： 570 nm
• 信号响应： 递增型

标准物质： NAD⁺ & NADH
参考标准： y=0.2525x-0.0059 (R²=0.9995)
标准线性范围： 0.25-5.0 nmol/mL
检测限： 0.1 nmol/mL

①试剂三和试剂四配制后有效期较短，为便于试验安排，各附赠一瓶作为备用，每瓶均可满足至少25个样本的测定；

②试剂五配制后未使用完试剂应置于4℃保存，严禁-20℃保存，使用过程中试剂五应用液需置于冰上放置；

③2 µmol/mL NAD⁺标准液和2 µmol/mL NADH标准液配制后4℃可保存1周，严禁-20℃保存；

④实验过程中吸取上清液时应避免吸入沉淀物而影响结果的准确性；

⑤反应时间对实验结果有较大影响建议精确控制，测定样本较多时建议分批进行测定，以避免组内反应时间不一致对结果造成影响；

⑥反应体系中试剂不能按比例配制为混合液使用，必须按反应体系顺序依次加入；

⑦若A测定大于1.0，建议将待测样本适当稀释后再进行测定，计算时相应修改；

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