Fluorescent Protein Fusion Vectors: mTFP1, mWasabi, FRET and Photoconvertible

Fluorescent Proteins Overview

With the 2008 Nobel Prize in Chemistry, the value of the discovery and development of fluorescent proteins (FPs) has been well recognized and appreciated. However, the research on FPs has not reached its logical end. Instead, there have been relentless endeavors to find the "perfect" FPs since the discovery of FPs just over a decade ago. Bright, photostable, monomeric, and fast-maturing FPs are being sought in all spectral regions. Those that work well as fusions or in fluorescent resonance energy transfer (FRET) see ever increasing use as molecular markers or rulers. Novel microscopy techniques that can image with resolutions of ~10

nm, previously only possible with electronic microscopy, rely on a family of FPs called photoactivitable or photoswitchable FPs. Such techniques include PALM for Photoactivated Localization Microscopy; SIM for Structure-Illumination Microscopy. Photoswitchable fluorescent proteins that emit larger number of photons before irreversible photobleaching and FPs that are significantly more photostable than current choices will be of great usefulness. Allele scientists work with other FP researchers with these photoswitchable fluorescent proteins to create new FP technologies and enable experiments not feasible otherwise.

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Choosing Fluorescent Proteins Despite the fact that many new or improved FPs recently reported are much more suitable for molecular imaging or other applications, most researchers still use the enhanced jellyfish GFP or its derivatives. Many users simply do not know about the better alternatives available to them or are deterred by the lack of commercial sources for easily obtaining reliable constructs...... Learn More	What Allele Offers Allele FP researchers and collaborators design, evolve, and select new FPs that suit different applications. What Allele now offers: mTFP1, a bright cyan FP with extremely high confocal photostability (2,721s) mWasabi, a green derivative of mTFP1 that is twice as bright as EGFP What´s in the pipeline: A green-to-red photoconvertible FP A family of new FPs from Branchiostoma (Amphioxus) in a wide spectral range
Properties of All^eleustrious mTFP1 Since its introduction in 2006 mTFP1 has been selected by hundreds of labs around the world and generated uniform positive feedback. mTFP1 is the brightest FP in the blue-cyan spectrum. With advantages such as being a true monomer, having desirable stability under even strong acidic conditions, mTFP1 has been tested and confirmed as a great fluorescent protein fusion tag to 24 different subcellular structure-specific polypeptides. It is also a top choice for donor in a FRET pair with orange or red FPs. Table of Properties	Properties of All^eleustrious mWasabi A recent addition to the FP toolbox, mWasabi is a top choice as a versatile fluorescent protein to be used as a general cellular marker and as a fusion tag for detection of proteins even at low expression levels in cells. As a favorable alternative to EGFP, mWasabi is available to commercial applications with simple licensing process and competitive terms. It is being integrated into other Allele products such as viral vectors or cell and animal models. Table of Properties
mTFP1 Is a Great FRET Donor Because of its excitation and emission wavelength, sharp excitation and emission peaks, high quantum yield, and exceptional photostability, mTFP1 has always been considered a very good Forster resonance energy transfer (FRET) donor (1). More recently, several groups have investigated the use of mTFP1 in various FRET experiments and imaging modalities and have shown that mTFP1 is indeed one of the best choices (2, 3, 4). (1) Ai et al., (2006) Biochem. J. 400:531-540. (2) Padilla-Parra et al., (2009) Biophys J. 97(8):2368-76. (3) Day et al., (2008) J Biomed Opt. 13(3):031203. (4) Sun et al., (2009) J Biomed Opt. 14(5):054009. Learn More	One-stop FRET Analysis Services Allele Biotech is one of few research groups that develop original FP technologies. Companies or academic labs will benefit from Allele Biotech’s vast expertise by engaging the Allele FP service, particularly in the more advanced fields such as FRET. The best FRET donor protein mTFP1 Option of integrating customer-supplied FPs into a FRET pair Full service cloning, cell culture, and viral packaging available if needed Mithras LB940 plate reader (FRET capable) and Biomek2000 robot for high throughput analysis Learn More