Blue Fluorescent Protein (BFP)
• Bright blue fluorescence
• Monomeric fast maturing protein with successful performance in fusions
• Extremely high pH-stability
• High photostability
• Recommended as a donor for green fluorescent proteins in FRET applications
TagBFP (scientific name mTagBFP) is a monomeric blue fluorescent protein generated by site-specific and random mutagenesis of TagRFP. TagBFP exhibits bright blue fluorescence with excitation/emission maxima at 402 and 457 nm, characterized by high photostability and extremely high pH-stability.
Compared to EBFP2, TagBFP is more then 1.8 times brighter, much more pH-stable and has twice shorter maturation half-time at 37°C. Narrow fluorescence emission peak of TagBFP provides for accurate and easy spectral separation with cyan and green fluorescent proteins and makes it a preferable tag for multicolor labeling.
Good overlap between the emission spectrum of TagBFP and the absorbance spectra of TagGFP allows using these two proteins as a FRET pair.
HeLa cells expressing TagBFP fusion proteins.
Performance and Use
TagBFP can be easily expressed and detected in a wide range of organisms. Mammalian cells transiently transfected with TagBFP expression vectors give bright fluorescent signals within 10-12 hrs after transfection. No cell toxic effects and visible protein aggregation are observed.
TagBFP performance in fusions has been demonstrated in the b-actin and a-tubulin models. It can be used in multicolor labeling applications with green, yellow, red, and far-red fluorescent dyes.
TagBFP can be successfully used as a donor in FRET pair also comprising TagGFP. The calculated Forster distance R0= 5.25 nm for TagBFP / TagGFP pair is larger than those reported for the standard ECFP-EYFP and mCyPet-mYPet pairs (R0= 4.86 nm and 4.93 nm correspondingly), suggesting that TagBFP / TagGFP is one of the best among available FRET pairs of monomeric fluorescent proteins. High efficiency of TagBFP as a FRET donor was demonstrated in living cells by Subach et al.
Recommended Filter Sets and Antibodies
The protein can be recognized using Anti-tRFP Antibody, see link below.
TagBFP can be detected using common fluorescence filter sets for BFP, DAPI, and other blue dyes.
Recommended filter sets are: XF119-2*, QMAX-Blue*, XF131, XF06, XF13-2, XF03, XF11, XF129-2, XF05-2 (Omega Optical); DAPI-5060B* and DAPI-1160A (Semrock); 31037, 31041, 31016*, 31021, 31000v2, 1009v2, 31013v2, 11005v2, 31047 (Chroma Technology Corp.).
* preferred filter sets
Subach OM et al. (2008) Conversion of Red Fluorescent Protein into a Bright Blue Probe. Chemistry & Biology 15 (10): 1116–1124.