NADP/NADPH 检测试剂盒 荧光法、NADP/NADPH Assay Kit *Fluorimetric*详细如下: NADP/NADPH 检测试剂盒利用优化的酶回收方法对 NADP/NADPH 进行定量,而不会干扰 NAD/NADH。NADP 在这种酶循环反应中转化为 NADPH。该试剂盒包含荧光试剂,与 NADPH 反应时产生荧光。产生的红色荧光可以在激发/发射= 560/590 nm 下进行监测。产生的荧光强度与 NADP/NADPH 浓度成正比。检测限:1.5 nM 烟酰胺核苷酸是细胞能量转化和氧化还原反应的关键参与者。NADP 和 NADPH 在调节酶活性中充当变构效应物。NADPH 的还原能力是促氧化剂酶(如一氧hua氮合酶和 NADPH 氧化酶)所需的辅助因子。NADP/NADPH 比率参与氧化分流的控制。这些形式中的每一种在分化和相似过程中都具有特定的作用,并且在电离辐射和烷化剂的细胞毒作用中具有目标功能。 Note: Avoid reducing agents (e.g. dithiothreitol, DTT; β-mercaptoethanol) in test samples. If samplescontain reducing agent, it should be neutralized by N-ethylmaleimide (NEM) using 2:1 NEM to DTT molar ratio. This assay tolerates the presence of sodium azide (up to 1 mM) and provides uncompromised signal in the presence of nonionic detergents such as Tween-20 (up to 10%), NP-40 (up to 1%) and Triton X-100 (up to 10%). Strong ionic detergents (e.g. SDS) completely inhibit assay. Collect cells by centrifugation at 2,500 rpm for 5 min and wash cell pellets by cold PBS. Extract cells with either 200 l of NADP extraction buffer (Component F) for NADP detection or NADPH extraction buffer (Component G) for NADPH detection. Add 200ul of assay buffer (Component D) Incubate cell suspensions at 60oC for 30 min. and then chill the samples on ice. Add 200 ul of opposite extraction buffer for neutralization. Quickly spin samples at 14,000 rpm for 5 min. Use supernatant for NADP/NADPH assay. Note: Typically, extract from 1x 105 – 5x105cells is used for one assay. Use several dilutions of sample to obtain readings that fit in the assay linear range.