HiLyte Fluor - Eurogentec

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2 Superior Fluorescent Labeling Dyes Spanning the Full Visible Spectrum HiLyte Fluor dyes, AnaSpec's patented dyes are a series of excellent fluorescent labeling dyes that span the full visible spectrum. For most of the HiLyte Fluor dyes, their hydrophilic property makes conjugations to protein easier, minimizing the use of organic solvents. The resulting conjugates are relatively resistant to precipitation during storage. The HiLyte Fluor dyes also have better labeling performance than the classic fluorescent labeling dyes such as FITC and rhodamines. Other features of the HiLyte Fluor dyes include: • HiLyte Fluor dyes are available in a variety of reactive forms. • HiLyte Fluor conjugates exhibit more intense fluorescence than other spectrally similar conjugates of classic fluorescent dyes such as FITC, TAMRA and ROX under similar excitations. • HiLyte Fluor dyes are more photostable than the classic fluorescent dyes such as FITC, Cy3 and Cy5. • HiLyte Fluor dyes have absorption spectra that match the principal output wavelengths of common excitation sources such as 488 nm, 555 nm, 633 nm and 647 nm. • HiLyte Fluor dyes and their conjugates are available in several distinct fluorescent colors. • HiLyte Fluor dyes are highly fluorescent over a broad pH range with little pH sensitivity. HiLyte Fluor dyes are the products of our R&D efforts. Additionally, AnaSpec is rapidly expanding our product lines to meet research needs. We have been developing dyes to solve various limitations with the existing fluorescent labeling reagents while offering classic fluorescent labeling reagents which have high purity and competitive pricing. We also provide custom synthesis and contract research services to meet your special needs. Our strong expertise in both the chemistry of fluorescent dyes and peptides equip us to provide quality services with competitive prices. 1. HiLyte Fluor 488 2. HiLyte Fluor 555 3. HiLyte Fluor 594 4. HiLyte Fluor 647 5. HiLyte Fluor 680 6. HiLyte Fluor 750 1. HiLyte Fluor 488 2. HiLyte Fluor 555 3. HiLyte Fluor 594 4. HiLyte Fluor 647 5. HiLyte Fluor 680 6. HiLyte Fluor 750 Normalized Absorbance Normalized Fluorescence 1.2 1.0 0.8 0.6 0.4 0.2 3 4 5 6 0.0 400 500 600 700 800 Wavelength (nm) 1.2 1.0 0.8 0.6 0.4 0.2 0.0 © 2009 AnaSpec, Inc. • 2149 O’Toole Ave. • San Jose, CA 95131 • 800-452-5530 • www.anaspec.com 1 1 2 2 3 4 5 6 500 600 700 800 900 Wavelength (nm) Figure 1. Absorption and emission spectra of AnaSpec's patented HiLyte Fluor dyes.
HiLyte Fluor 488 Dye, an Excellent Replacement for FITC Although FITC is still the most popular fluorescent labeling dye for preparing green fluorescent bioconjugates, there are limitations, such as severe photobleaching for microscope imaging and pH-sensitive fluorescence. Protein conjugates prepared with HiLyte Fluor 488 dyes (Ex/Em=497 nm/525 nm) are far superior compared to conjugates of fluorescein derivatives such as FITC (Figure 2). HiLyte Fluor 488 conjugates are significantly brighter than fluorescein conjugates and are much more photostable. Extinction coefficient is 70,000 M -1 cm -1 , with a quantum yield of 0.91. Additionally, the fluorescence of HiLyte Fluor 488 is not affected by pH (4-10). This pH insensitivity is a major improvement over fluorescein, which emits its maximum fluorescence only at pH above 9. Absorbance 1.2 1.0 0.8 0.6 0.4 Product Size Cat# HiLyte Fluor 488 acid 10 mg 81160 HiLyte Fluor 488 amine, TFA salt 1 mg 81162 HiLyte Fluor 488 C2 maleimide 1 mg 81164 HiLyte Fluor 488 hydrazide 1 mg 81163 HiLyte Fluor 488 succinimidyl ester (SE) 1 mg 5 mg 81161-1 81161 Normalized Absorbance 1.2 1.0 0.8 0.6 0.4 0.2 0.0 HiLyte Fluor 488 FITC A 0.0 400 500 600 700 Wavelength (nm) B C % Original Fluorescence 94 0 20 40 60 80 100 120 0 50 100 150 200 Irradiation Time (min) Time (sec) Figure 3. Tubulin in human glioblastoma cells was probed with mouse anti-tubulin, visualized with HiLyte Fluor 488-conjugated rabbit anti-mouse IgG (Cat# 28164-H488). Nuclei were stained with Hoechst 33342 (Cat# 83218). 106 104 102 100 98 96 HiLyte Fluor 488 Figure 2. Absorption and emission spectra of HiLyte Fluor 488 (panel A). Panels B and C show the photostability of HiLyte Fluor 488 compared to FITC and FAM. © 2009 AnaSpec, Inc. • 2149 O’Toole Ave. • San Jose, CA 95131 • 800-452-5530 • www.anaspec.com 1.2 1.0 0.8 0.6 0.4 0.2 Normalized Fluorescence 5-FAM FITC 3
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