dsRFP detection

Beat Ludin bludin at lis.ch
Thu Feb 17 06:41:56 EST 2000

To all of those who have contributed (or are planning to contribute) 
to this thread:

In view of the fact you seem to get different results concerning the 
brightness of DsRed compared to other FPs, would you mind describing 
your imaging setup (excitation source, filters, detector/camera 
model)? This would help a lot to get a better picture of the 



>Jered Floyd wrote:
>  > MMILLING at MEDAU.JNJ.com ("MILLINGTON, Michelle [JJRAU]") writes:
>  > > We have tried EBFP but are unable to detect by flow due to  low 
>  > > and photobleaching.  Is dsRFP brighter and more resistant to 
>  > > than EBFP?
>  >
>  > With an epifluorescence microscope, DsRed is significantly brighter
>  > and much more resistant to photobleaching than GFP/EGFP;
>We´ve also used RFP and EGFP, but with reverse result: EGFP is MUCH 
>brighter the
>RFP, at least in COS cells transfected with vectors dsRed-N1 and pEGFP-N1,
>  > I imagine as
>  > such it is much better than EBFP as well.  I have not yet tried it
>  > with flow, but will soon.
>  >
>  > One word of warning with DsRed: At least in E. coli, there is a very
>  > large delay (36-48 hours) before it is visible to the eye. This is
>  > unfortunate, as the other FPs express visibly much faster.
>  >
>Not only in E.coli. When looking at transfected cells, RFP is not 
>detectable 24h
>after transfection (when EGFP is already VERY bright), but shows a good signal
>only after at least 48h. Seems the "maturation" of the protein - 
>which makes it
>fluorescent - takes much longer for RFP than for GFP. This is an 
>important thing
>to remember when doing cotransfections with RFP and GFP.

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