Plant Molecular Biology Reporter

, Volume 25, Issue 1–2, pp 45–54 | Cite as

A Rapid and Sensitive Fluorimetric Protocol for the Quantification of Green Fluorescent Protein in Soluble Protein Extracts from Transgenic Arabidopsis

  • Wenche Johansen
  • Else-Berit Stenseth
  • Robert C. Wilson


Green fluorescent protein (GFP) is a popular qualitative reporter protein used to study different aspects of plant biology. However, to be used as a reliable quantitative reporter in expression studies using fluorescence based assays, methods to eliminate interfering endogenous molecules must be considered. Therefore, a standard curve based solid phase fluorescent immunoassay that eliminates the effects of interfering endogenous molecules was developed to quantify the GFP levels in soluble green extracts prepared from plants. Microtiter plates coated with anti-GFP were used to capture GFP from soluble plant extracts, interfering endogenous molecules was eliminated by washing without disturbing the anti-GFP binding of GFP, and then the fluorescence intensity of bound GFP was measured using a spectrofluorometer. We report in this study the use of this method to quantify the expression levels of soluble modified GFP in transgenic Arabidopsis thaliana.


Arabidopsis GFP quantification Solid-phase fluorescent immunoassay Soluble plant extract Transgene expression 



cauliflower mosaic virus


soluble modified green fluorescent protein


relative fluorescence units



We thank Dr. Phil Mullineaux and Dr. Roger Hellens at John Innes Centre for providing the pGII0029 vector and the Nottingham Arabidopsis Stock Centre (NASC) for providing the 35S-smGFP clone. We thank Johan R. Lillehaug, Department of Molecular Biology, University of Bergen, for helpful discussions. This work was supported by grants from the Hedmark County in Norway.


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Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Wenche Johansen
    • 1
  • Else-Berit Stenseth
    • 1
  • Robert C. Wilson
    • 1
  1. 1.Department of Natural Science and Technology, Faculty of Education and Natural ScienceHedmark University CollegeHamarNorway

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