Fluorescence Assay for Phospholipase C Activity Using Liposome Probes
10.11895/j.issn.0253-3820.170179
- VernacularTitle:脂质体荧光探针检测磷脂酶C的活性
- Author:
Qiaorong GU
;
Junjie AI
;
Qianyun ZHANG
;
Yanan DONG
;
Qiang GAO
- Keywords:
Fluorescence;
Liposome;
Probe;
Phospholipase C;
Inhibitor
- From:
Chinese Journal of Analytical Chemistry
2017;45(9):1278-1283
- CountryChina
- Language:Chinese
-
Abstract:
A simple assay for detection of phospholipase C (PLC) activity was developed based on a fluorescence liposome probe using the Liss Rhod PE-loaded phospholipid liposomes.The liposome probe was prepared by the coassembly of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and fluorescent lipid (Liss Rhod PE).The probe showed very low background fluorescence due to fluorescence self-quenching effect of Liss Rhod PE.As the PLC enzyme selectively digested lipid, the Rhod fluorescence was recovered from its quenched state, leading to the sensitive detection of PLC.This assay provided a limit of detection (at a signal-to-noise ratio of 3) of 2 U/L for PLC.In the presence of PLC inhibitor, the fluorescent response of the sensor for PLC decreased, indicating that the assay could also be used for screening PLC inhibitors.