Red/Far-red Protein Tags

[Michael Jacobsen-3]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hello,

 

This is a first-time post to this listserv.

 

Does anyone know of a red-excitable expression tag?

 

I am working with a 488/568/647 Ar and Ar/Kr laser confocal system, and I am looking for a protein tag that can be excited using a 647 laser. Ultimately, I would like to express proteins with three different tags: GFP(488), RFP(568), and ???(647).

 

Recently, I saw a discussion on this listserv about far-red tags mKate and JRed--but these are not "far" enough. I have also skimmed some of Tsien lab's recent works, but have not found any 600-excitable tags.

 

Any help or insight would be appreciated.

 

Thank you very much,

 

Michael Jacobsen

Manager, Confocal Microscopy Core Facility
Department of Pharmacology and Experimental Neuroscience
University of Nebraska, Medical Center

 

[Rosemary.White]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear Michael,

You may find that expression tags that have peak EX in far-red have sufficient EX at shorter wavelengths that you don’t need to worry about this.  For example, we use DsRed, Texas Red, mitotracker red and propidium iodide, all of which have EX peaks in the yellow-orange region, and I never bother using anything other than 488 nm EX, usually because we’re looking at GFP and autofluorescence at the same time.  Have not noticed either detection or bleaching problems.  Perhaps you’d have to worry with low-expressed probes, but we’ve not had trouble yet (cross fingers, etc....).

Might be worth a try, esp if you already have ready access to these other tags.

cheers,
Rosemary

Rosemary White                    [hidden email]
CSIRO Plant Industry            ph.     61 (0)2-6246 5475
GPO Box 1600                       fax.     [phone]
Canberra, ACT 2601            
Australia



On 14/11/07 6:09 AM, "Michael T Jacobsen" <[hidden email]> wrote:

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hello,

 

This is a first-time post to this listserv.

 

Does anyone know of a red-excitable expression tag?

 

I am working with a 488/568/647 Ar and Ar/Kr laser confocal system, and I am looking for a protein tag that can be excited using a 647 laser. Ultimately, I would like to express proteins with three different tags: GFP(488), RFP(568), and ???(647).

 

Recently, I saw a discussion on this listserv about far-red tags mKate and JRed--but these are not "far" enough. I have also skimmed some of Tsien lab's recent works, but have not found any 600-excitable tags.

 

Any help or insight would be appreciated.

 

Thank you very much,

 

Michael Jacobsen

Manager, Confocal Microscopy Core Facility
Department of Pharmacology and Experimental Neuroscience
University of Nebraska, Medical Center