TagYFP

Yellow fluorescent protein TagYFP

- Bright yellow fluorescence
- Monomeric protein with successful performance in fusions
- Fast maturation, high pH-stability and photostability
- Proven suitability to generate stably transfected cell lines
- Recommended for protein labeling

TagYFP is a monomeric yellow fluorescent protein developed on the basis of GFP-like protein from jellyfish Aequorea macrodactyla [Xia et al., 2002]. TagYFP possesses single excitation maximum at 508 nm, and emission maximum at 524 nm. TagYFP is more pH stable than EYFP.

TagYFP is mainly intended for protein labeling in protein localization and interaction studies. It can also be used for cell and organelle labeling and for tracking the promoter activity, although TurboYFP and Phi-Yellow proteins are preferable for such applications because they mature faster and give brighter fluorescent signal.

Main properties

TagYFP spectra

TagYFP normalized excitation (thin line) and emission (thick line) spectra.

Spectra viewer tool
Download TagYFP spectra (xls)

CHARACTERISTIC
* Brightness is a product of extinction coefficient and quantum yield, divided by 1000.
Molecular weight, kDa27.0
Polypeptide length, aa239
Fluorescence coloryellow
Excitation maximum, nm508
Emission maximum, nm524
Quantum yield0.62
Extinction coefficient, M-1cm-150 000
Brightness*31
Brightness, % of EGFP94
pKa5.5
Structuremonomer
Aggregationno
Maturation rate at 37°Cfast
Photostabilityhigh
Cell toxicitynot observed
Main advantagesbright yellow monomeric fluorescent protein

Recommended filter sets and antibodies

The protein can be recognized using Anti-Tag(CGY)FP antibody (Cat.# AB121-AB122) available from Evrogen.

Recommended Omega Optical filter sets for TagYFP are XF104-3 and XF105-2. It can also be detected using Chroma Technology Corp. filter set 41028 Yellow GFP BP (10C/Topaz) or the similar.

Performance and use

TagYFP can be easily expressed and detected in a wide range of organisms. Mammalian cells transiently transfected with TagYFP expression vectors produce bright fluorescence in 10-12 hrs after transfection. No cytotoxic effects or visible protein aggregation are observed.

TagYFP use for labeling of cells and cellular orgenelles.

Confocal microscopy of transiently transfected mammalian cells expressing TagYFP.

TagYFP performance in fusions has been demonstrated in human cytoplasmic β-actin and α-tubulin models. An expected pattern of fluorescence has been obtained in each case.

TagYFP use for protein labeling.

Confocal microscopy of transiently transfected mammalian cells expressing TagYFP fusions.

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

Stably transfected cell lines expressing TagYFP fusions.

(A) MDCK cells expressing TagYFP-tubulin fusion; (B) U-205 cells expressing TagYFP-actin fusion. Images were kindly provided by Dr. Christian Petzelt (Marinpharm).

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

Multicolor labeling using TagFPs.

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


Available variants and fusions
VariantDescriptionRelated vectorCat.#Click for image
Humanized TagYFP TagYFP codon usage is optimized for high expression in mammalian cells [Haas et al., 1996], but it can be successfully expressed in many other heterological systems. pTagYFP-C FP131
pTagYFP-N FP132
TagYFP-actin fusion Human β-actin is fused to the TagYFP C-terminus. When expressed in mammalian cells, this fusion provides yellow fluorescent labeling of β-actin in living cells. pTagYFP-actin FP134
TagYFP-tubulin fusion Human α-tubulin is fused to the TagYFP C-terminus. When expressed in mammalian cells, this fusion provides yellow fluorescent labeling of α-tubulin in living cells. pTagYFP-tubulin FP135
TagYFP-mito fusion A mitochondrial targeting sequence (MTS) is fused to the TagYFP N-terminus. MTS was derived from the subunit VIII of human cytochrome C oxidase [Rizzuto et al., 1989; Rizzuto et al., 1995]. When expressed in mammalian cells, this variant provides yellow fluorescent labeling of mitochondria. pTagYFP-mito FP137


References:

  • Haas J, Park EC, Seed B. Codon usage limitation in the expression of HIV-1 envelope glycoprotein. Curr Biol. 1996; 6 (3):315-24. / pmid: 8805248
  • Rizzuto R, Brini M, Pizzo P, Murgia M, Pozzan T. Chimeric green fluorescent protein as a tool for visualizing subcellular organelles in living cells. Curr Biol. 1995; 5 (6):635-42. / pmid: 7552174
  • Rizzuto R, Nakase H, Darras B, Francke U, Fabrizi GM, Mengel T, Walsh F, Kadenbach B, DiMauro S, Schon EA. A gene specifying subunit VIII of human cytochrome c oxidase is localized to chromosome 11 and is expressed in both muscle and non-muscle tissues. J Biol Chem. 1989; 264 (18):10595-600. / pmid: 2543673
  • Xia NS, Luo WX, Zhang J, Xie XY, Yang HJ, Li SW, Chen M, Ng MH. Bioluminescence of Aequorea macrodactyla, a common jellyfish species in the East China Sea. Mar Biotechnol (NY). 2002; 4 (2):155-62. / pmid: 14961275
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