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Green Fluorescent Protein (GFP)

Ursa BioScience offers a highly pure form of Green Fluorescent Protein (GFP) from Aequorea Victoria (Recombinant wt GFP), purified with high resolution gel-filtration as the final step.

Green fluorescent protein (GFP), a 27 kDa protein derived from the jellyfish Aequorea victoria, emits green light (emission peak at a wavelength of 509 nm) when excited by blue light (excitation peak at a wavelength of 395 nm). Green Fluorescent Protein (GFP) has become an invaluable tool in cell biology research [1,2], since its intrinsic fluorescence can be visualized in living cells. GFP fluorescence is stable under fixation conditions and suitable for a variety of applications. GFP has been widely used as a reporter for gene expression, enabling researchers to visualize and localize GFP-tagged proteins within living cells without the need for chemical staining. Other applications of GFP include assessment of protein-protein interactions through the yeast two hybrid system and measurement of distance between proteins through fluorescence energy transfer (FRET) protocols. GFP is used to measure single cell metastasis and successful proliferation of stem cells. In these ways, GFP technology has contributed to a greater understanding of cellular biology and biochemistry.

Figure 1A shows the 3D Excitation - Emission Matrix (EEM) recorded for wt-GFP in Tris-EDTA, pH 7.6 at 21 C. The white circles indicates where fluorescence lifetimes have been recorded, see Table I. Figure 1B shows the anisotropy contour map corresponding to data shown in Figure 1A. Table 1 shows the fluorescence lifetimes of the wt-GFP dissolved in Tris-EDTA, pH 7.6, 21 C.

Jelly fish

Figure 1. (A) Excitation-Emission Map (EEM) recorded for WT-GFP dissolved in Tris-EDTA (pH 7.6)  at + 21 C. The circles (white) indicates where fluorescence lifetimes were recorded, see Table 1. The color map used for the intensity scale is set to 1 at the peak centered around (510,390) nm. (B) The anisotropy map corresponding to data in Panel A. Note that the contour lines are shown for the intensity data, and not for the anisotropy values.

 

Figure 2 shows the thermal stability of wt-GFP in Tris-EDTA, pH 7.6, during a heat and cool cycle. The excitation wavelength was 460 nm. The fluorescence signal is the integrated emission band centered at 500 nm. The sample is firstly heated from +5 C to +60 C, then the sample is cooled back to + 5 C again.

Heat stability GFP

Figure 2. Fluorescence response from WT-GFP, dissolved in Tris-EDTA (pH 7.6), during a heat and cool cycle. The excitation wavelength was 460 nm. The fluorescence signal is the integrated emission band centered at 500 nm. First the sample is heated from +5 C to +60 C, then the sample is cooled back to + 5 C again.

 

The purity of our GFP, from an E. Coli host, is confirmed by constant specific absorbance across chromatographic peaks, SEC-HPLC, and A397/A280 ratios.

Cited References

[1] Chalfie, M., Y. Tu, G. Euskirchen, W.W. Ward and D.C. Prasher. 1994. Green-fluorescent protein as a marker for gene expression. Science 263:802-805.
[2] Prasher, D.C., V.K. Eckenrode, W.W. Ward, F.G. Prendergast, and M.J. Cormier. 1992. Primary structure of the Aequorea victoria greenfluorescent protein. Gene 111:229-233.

 

 

Green fluorescent protein

Structure of Green Fluorescent Protein (GFP).

 

Jelly 4

Jelly 3

 Aequorea Victoria