Flow Cytometric Analysis of GFP Expression in Mammalian Cells

  • Steven R. Kain
Chapter
Part of the Springer Lab Manuals book series (SLM)

Abstract

In the jellyfish Aequorea victoria, light is produced when energy is transferred from the Ca2+-activated photo protein aequorin to green fluorescent protein or GFP (Fig. 1).1-3 This process occurs in specialized photogenic cells located at the base of jellyfish umbrella, where each protein is found at very high concentrations. The cloning of the wild-type GFP gene (wt GFP)4,5 and its subsequent expression in heterologous systems6,7 has established GFP an important reporter protein for the analysis of gene expression and protein localization in a wide variety of experimental designs. When expressed in either eukaryotic or prokaryotic cells and illuminated by blue or UV light, wt GFP emits a bright green fluorescent signal which is easily detected by fluorescence microscopy, flow cytometry, or other fluorescence imaging techniques. GFP fluorescence is species-independent and does not require additional cofactors, substrates, or gene products — the protein is “naturally fluorescent”. Moreover, detection of GFP and its variants can be performed in living samples, and is ideally suited to real time analysis of molecular events.

Keywords

Green Fluorescent Protein Flow Cytometric Analysis Green Fluorescent Protein Expression Green Fluorescent Protein Gene Green Fluorescent Protein Variant 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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© Springer-Verlag Berlin Heidelberg 2000

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  • Steven R. Kain

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