TagRFP

Red fluorescent protein TagRFP

- Bright red (orange) fluorescence
- Monomeric protein with successful performance in fusions
- Fast maturation, high pH-stability
- Proven suitability to generate stably transfected cell lines
- Recommended for protein labeling, acidic organelle labeling, FRET applications

Performance and use

TagRFP can be easily expressed and detected in a wide range of organisms. Mammalian cells transiently transfected with TagRFP expression vectors produce bright fluorescence in 10-12 hrs after transfection. No cytotoxic effects or visible protein aggregation are observed. High pH-stability with pKa=3.8 makes it possible to use TagRFP for imaging in acidic organelles, such as late and recycling endosomes and lysosomes.

TagRFP use for cell and organelle labeling.

Microscopic images of HeLa cells transiently transfected with TagRFP and TagRFP-targeted to cellular organelles. Images were kindly provided by Michael W. Davidson (Florida State University).

TagRFP performance in protein fusions has been demonstrated in fibrillarin, vinculin, zyxin, β-actin, α-tubulin, and other models.

TagRFP use for protein labeling in mammalian cells.

Microscopic images of HeLa cells transiently transfected with TagRFP-tagged fusions. Images were kindly provided by Michael W. Davidson (Florida State University).

TagRFP suitability to generate stably transfected cells has been proven by Marinpharm company. Cell lines expressing TagRFP are commercially available.

Stably transfected cell lines expressing TagRFP fusions.

U-205 mammalian cell lines expressing (A) TagRFP fusion with α-tubulin and (B) TagRFP fusion with β-actin. Images were kindly provided by Dr. Christian Petzelt (Marinpharm).

TagRFP can be used in multicolor labeling applications with blue, cyan, green, yellow, and far-red fluorescent dyes.

TagRFP use in multicolor labeling of mammalian cells.

(A) TagCFP-tagged β-actin (cyan), TagYFP-tagged α-tubulin (yellow), TagFP635-H2B fusion (red), and Golgi-targeted TagRFP (violet); (B) mitochondria-targeted TagCFP (cyan), Dendra2-tagged vimentin (green), and TagRFP-tagged β-actin (red). Image (A) was kindly provided by Michael W. Davidson (Florida State University).

TagRFP is ideally suitable for use as an acceptor for FRET with Evrogen green fluorescent protein TagGFP2. The calculated Forster distance (R0 = 5.7 nm) for the TagGFP2-TagRFP pair is one of the largest among the values reported. At the same time, since TagGFP2 and TagRFP emission peaks are spaced by as much as 78 nm, the emission signal for these two proteins can be easily separated in any imaging system. High pH-stability of the both proteins allows using this pair for imaging in acidic organelles.

The excellent performance of TagRFP in FRET application was demonstrated both in vitro and in vivo on the example of FRET-based apoptosis reporter Casper3-GR [Shcherbo et al., 2009].

References:

  • Shcherbo D, Souslova EA, Goedhart J, Chepurnykh TV, Gaintzeva A, Shemiakina II, Gadella TW, Lukyanov S, Chudakov DM. Practical and reliable FRET/FLIM pair of fluorescent proteins. BMC Biotechnol. 2009; 9 :24. doi: 10.1186/1472-6750-9-24 / pmid: 19321010
Copyright 2002-2016 Evrogen. All rights reserved.
Evrogen JSC, 16/10 Miklukho-Maklaya str., Moscow, Russia, Tel +7(495)988-4084, Fax +7(495)988-4085, e-mail:evrogen@evrogen.com