A. Knight, N. Goddard, P. Fielden, M. Barker, N. Billinton, R. Walmsley
{"title":"绿色荧光蛋白(GFP)的荧光偏振。一种改进GFP测定波长辨别的策略","authors":"A. Knight, N. Goddard, P. Fielden, M. Barker, N. Billinton, R. Walmsley","doi":"10.1039/A901326A","DOIUrl":null,"url":null,"abstract":"The fluorescence of green fluorescent protein (GFP), present both within whole yeast cells and in protein extracts from yeast cells, has been observed to be significantly polarised. Fluorescence polarisation is proposed as a useful technique to allow some discrimination between GFP fluorescence and that of other interfering species in cell or media matrices, which have fluorescence bands that overlap those of GFP, which should lead to improved resolution and limits of detection. The method has been tested by discriminating between the fluorescence of GFP in cell extracts and added fluorescein, both of which fluoresce brightly at the same wavelength. The flow-through instrumentation incorporating an argon-ion laser developed for this work is also described.","PeriodicalId":7814,"journal":{"name":"Analytical Communications","volume":"27 1","pages":"113-117"},"PeriodicalIF":0.0000,"publicationDate":"1999-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"14","resultStr":"{\"title\":\"Fluorescence polarisation of green fluorescent protein (GFP). A strategy for improved wavelength discrimination for GFP determinations\",\"authors\":\"A. Knight, N. Goddard, P. Fielden, M. Barker, N. Billinton, R. Walmsley\",\"doi\":\"10.1039/A901326A\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"The fluorescence of green fluorescent protein (GFP), present both within whole yeast cells and in protein extracts from yeast cells, has been observed to be significantly polarised. Fluorescence polarisation is proposed as a useful technique to allow some discrimination between GFP fluorescence and that of other interfering species in cell or media matrices, which have fluorescence bands that overlap those of GFP, which should lead to improved resolution and limits of detection. The method has been tested by discriminating between the fluorescence of GFP in cell extracts and added fluorescein, both of which fluoresce brightly at the same wavelength. The flow-through instrumentation incorporating an argon-ion laser developed for this work is also described.\",\"PeriodicalId\":7814,\"journal\":{\"name\":\"Analytical Communications\",\"volume\":\"27 1\",\"pages\":\"113-117\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1999-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"14\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Analytical Communications\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1039/A901326A\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Analytical Communications","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1039/A901326A","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Fluorescence polarisation of green fluorescent protein (GFP). A strategy for improved wavelength discrimination for GFP determinations
The fluorescence of green fluorescent protein (GFP), present both within whole yeast cells and in protein extracts from yeast cells, has been observed to be significantly polarised. Fluorescence polarisation is proposed as a useful technique to allow some discrimination between GFP fluorescence and that of other interfering species in cell or media matrices, which have fluorescence bands that overlap those of GFP, which should lead to improved resolution and limits of detection. The method has been tested by discriminating between the fluorescence of GFP in cell extracts and added fluorescein, both of which fluoresce brightly at the same wavelength. The flow-through instrumentation incorporating an argon-ion laser developed for this work is also described.