Q:P450试剂盒的选择
P450-Glo™ 检测系统为测定细胞色素P450活性提供了一种均质的发光检测方法。该检测系统使用的P450发光前体底物是甲虫萤光素（beetle luciferin）的衍生物，甲虫萤光素是萤光素酶的底物。此衍生物本身不是萤光素酶的底物，但是当P450酶将其转化为萤光素时，即成为萤光素酶底物，并与萤光素酶反应而产生光，光强度与P450活性成正比。
P450-Glo™ 检测系统可以用来检测：
- CYP抑制：筛选对天然或重组组分中的CYP活性有抑制作用的药物和新的化合物。
- 重组CYP活性：测定来自昆虫细胞和大肠杆菌等外源表达系统的膜部分的重组CYP活性。
- 天然CYP活性：测定来自组织(如肝脏)的微粒体等部分的天然CYP活性。
- CYP诱导：通过测量完整细胞(如肝细胞)内的CYP活性来识别和鉴定CYP基因表达的诱导因子。
Promega同时提供P450-Glo™ Screening System，用于P450相关抑制剂或药物的筛选，系统中包括含有通过膜制备的重组人细胞色素P450（CYP）酶，阴性对照膜，适合CYP酶的发光底物，NADPH再生系统，反应缓冲液，萤光素酶检测试剂和无萤光素水。系统经过优化后，可以快速稳定的进行CYP酶类抑制剂等化合物的高灵敏度筛选。
|
底物种类 |
P450酶活检测（生化检测） |
细胞水平检测 |
|
Luciferin-ME |
CYP1A2, CYP2C8, CYP2C9, CYP2J2, CYP4A11, CYP4F3B, CYP19 |
Not recommended |
|
Luciferin-1A2 |
CYP1A2 |
CYP1A2 induction |
|
Luciferin-CEE |
CYP1A1, CYP1B1, CYP3A7 |
CYP1A induction |
|
Luciferin-2B6 |
CYP2B6 |
CYP2B6 induction |
|
Luciferin-H |
CYP2C9 |
CYP2C9 induction |
|
Luciferin-ME EGE |
CYP2D6, CYP1A1, CYP1A2, CYP2B6 |
Not recommended |
|
Luciferin-H EGE |
CYP2C19, CYP1A1, CYP1A2 |
Not recommended |
|
Luciferin-BE |
CYP3A4, CYP3A5, CYP3A7, CYP4F12 |
Not recommended |
|
Luciferin-PFBE |
CYP3A4, CYP3A5, CYP3A7 |
CYP3A induction |
|
Luciferin-PPXE |
CYP3A4, CYP3A5, CYP3A7 |
Not recommended |
|
Luciferin-IPA |
CYP3A4 |
CYP3A induction |
This assay can be used to measure CYP1A2 activity in hepatocytes. Luciferin-1A2 is highly selective for CYP1A2.
This assay can be used to measure CYP2B6 activity in hepatocytes. Luciferin-2B6 is highly selective for CYP2B6.
This assay can be used to measure CYP2C9 activity in human hepatocytes and other human cell types. Luciferin-H is highly selective for CYP2C9.
The P450-Glo™ CYP1A1 Assay can be used to selectively measure CYP1A1 activity in hepatocytes where cross-reacting CYP1B1 and CYP3A7 are absent.
