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|---|---|---|---|
| 100 ul | $303.00 | Add to Cart |
The green fluorescent protein (GFP) is a 27kDa protein isolated originally from the jellyfish Aequoria victoria. It has an endogenous fluorochrome activity with excitation maximum at 395nm and emission maximum at 509nm, which is similar to that of fluorescein. The GFP gene was sequenced and the origin of the fluorochrome by autocatalytic activity of certain amino acids was discovered. Much interest in GFP was generated when it was shown that fluorescence develops rapidly when the protein is expressed and requires only molecular oxygen and no other cofactors. As a result GFP can be expressed in fluorescent form in essentially any prokaryotic or eukaryotic cell. GFP has been engineered to produce a vast number of variously colored mutants including blue, cyan and yellow protein derivatives, BFP, CFP and YFP. GFP and other fluorescent proteins derived from other Cnidarians (jellyfish, coral and medusa) are widely used as tracers in transfection and transgenic experiments to monitor gene expression and protein localization in vivo and in in vitro. The crystal structure of GFP was determined which allowed amino acid modifications to improve spectral properties and prevent multimerization. |
Images
Immunofluorescent analysis of transfected HEK293 cells transfected with a GFP construct and stained with mouse mAb to GFP, MO22190, dilution 1:1,000, in red. The blue is Hoechst staining of nuclear DNA. The GFP antibody binds to GFP protein expressed only in transfected cells, and as a result cells are appeared in orange-golden color.
Western blot analysis of transfected and control HEK293 cell lysates using mouse mAb to GFP, MO22190, in green, dilution 1:1,000: [1] protein standard, [2] Control, non-transfected cells, [3] cells transfected with an mCherry red fluorescent protein construct and [4] cells transfected with GFP construct. The strong green band at ~27kDa corresponds to GFP protein detected only in cells transfected with GFP construct, the antibody does not bind to mCherry.