EGFP and GFP - how different are they? (Sep/02/2006 )

hi
how different are EGFP and GFP from each other?
and does GFP antibody recognise EGFP also? if they both require different antibody can tell me their names
thanks

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The EGFP (Enhanced GFP) and GFP differ for a few aminoacids that make EGFP fluoresce stronger than GFP. An anti-GFP antibody can recognize also EGFP. I use an anti-GFP from Abcam to recognize EGFP.

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EGFP is a mutant of GFP. It has increased fluorescence and the protein major excitation peak has been shifted to 490 nm with the peak emission kept at 509 nm (EGFP).

U should b able to detect EGFP with anti-GFP antibody.

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EGFP is also thought to be a bit less toxic to the cells expressing. All GFP antibodies I've worked with identify EGFP as well as native GFP. The one from clontech is great!

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Is it possible for changing EGFP in to RFP or YFP by simple mutagenesis?

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polyclonal anti-GFP of Cell signaling works well, no difference for original GFP, EGFP or mGFP;
CFP and YFP are mutations of GFP, I think cloning with the corresponding vectors appears to be more simply than mutating GFP; RFP? - never heard of it, DSred (Clontech) works fine

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I would prefer to work with the respective fluorescent protien vectors than mutating them. Its much easier. Instead of YFP, I would work with venus ( a modified version of YFP) and for RFP, I would work with DsRed2 or mCherry or mStrawberry (Tsien's lab)- depending on the experiments.

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dsRed and the other red fluorescent proteins are quite distinct from the GFPs, so antibodies to GFP will not function with them. The CFP, YFP and eGFP variants are all small changes from GFP and will be detected by GFP antibodies.

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I've heard that these are different because of a point mutation. Phenylalanine to Leucine if I remember right.

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