Yellow fluorescent protein TagYFP

Abstract

-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 of TagYFP Characteristic Molecular weight, kDa 27.0 Polypeptide length, aa 239 Fluorescence color yellow Excitation maximum, nm 508 Emission maximum, nm 524 Quantum yield 0.62 Extinction coefficient, M -1 cm -1 50 000 Brightness* 31 Brightness, % of EGFP 94 pKa 5.5 Structure monomer Aggregation no Maturation rate at 37°C fast Photostability high Cell toxicity not observed * Brightness is a product of extinction coefficient and quantum yield, divided by 1 000. Performance and use TagYFP can be easily expressed and detected in a wide range of organisms. Mammalian cells transiently transfected with TagYFP expression vectors give bright fluorescent signals in 10-12 hrs after transfection. No cytotoxic effects or visible protein aggregation are observed. 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 suitability to generate stably transfected cells has been proven by Marinpharm company. Cell lines expressing TagYFP are commercially available. TagYFP can be used in multicolor labeling applications with blue, cyan, red, and far-red fluores-cent dyes. 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. 0 20 40 60 80 100 Fluorescence, % 300 400 500 600 700 800 Wavelength, nm TagYFP normalized excitation (thin line) and emission (thick line) spectra. TagYFP expression in mammalian cells. (A) Confocal microscopy of cytoplasmic TagYFP expres-sion in transiently transfected human HeLa cells; (B) confocal microscopy of mitochondria-targeted TagYFP expression in transiently transfected HeLa cells; (C) con-focal microscopy of TagYFP fusion with the cytoplasmic β-actin in transiently transfected 3T3 cells; (D) confo-cal microscopy of TagYFP fusion with the α-tubulin in transiently transfected 3T3 cells.