Innovative Tools for Molecular and Cell Biology

Innovative Tools for Molecular and Cell Biology


Fluorescent Dyes

HiLyte™ Fluor Dyes


Absorption and Emission Spectra of AnaSpec's HiLyte Fluor™ Dyes

HiLyte Fluor 405

Alpha-tubulin in 3T3 cells probed with mouse anti-alpha tubulin antibody, visualized with HiLyte™ Fluor 405 conjugated goat anti-mouse IgG.

HiLyte Fluor 488

Tubulin in human glioblastoma cells was probed with mouse anti-tubulin, visualized with HiLyte™ Fluor 488 conjugated rabbit anti-mouse IgG. Nuclei were stained with Hoechst 33342 (Cat# 83218AS).

HiLyte Fluor 555

3T3 cells incubated with anti-tubulin antibody and goat anti-rabbit antibodies, labeled with HiLyte™ Fluor 555. Nuclei were stained with Hoechst 33342 (Cat# 83218AS).

HiLyte Fluor 750

CHO-K1 cells labeled with rabbit anti-tubulin primary antibody and a secondary antibody goat anti-rabbit IgG conjugated to HiLyte Fluor™ 750 (red); Paxillin EGFP (green) and a nuclear stain, DAPI (blue). Image courtesy of Kimberly Young and Claire M. Brown, McGill University Imaging Facility, Montreal, Canada.



  • 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
  • More photostable than the classic fluorescent dyes
  • Highly fluorescent over a broad pH range (4-11) with little pH sensitivity
  • Well-separated spectra, enabling many options for multicolor detection of multiple parameters simultaneously in flow cytometry analysis
  • Can be paired with proprietary QXL Quenchers for FRET
  • Cost-effective


  • AnaSpec's proprietary pH-insensitive HiLyte™ Fluor dyes are a series of excellent fluorescent labeling dyes with fluorescence emissions that span the full visible and near-infrared spectrum:

HiLyte™ Fluor 405

  • Available in a variety of reactive forms, such as amine-reactive, thiol-reactive, and amine-containing derivatives, allowing easy conjugation to biopolymers.
  • Spectral characteristics similar to those of Alexa Fluor™ 405 (Invitrogen) and DyLight Fluor™ 405 (Pierce) dyes with excitation and emission wavelength at 404/428 nm in PBS buffer (pH 7.4).
  • Conjugates of HiLyte™ Fluor 405 are well excited by the 407 nm spectral line of the Krypton laser and the 408 nm line of the violet laser diode.
  • Ex/Em = 404/428 nm, extinction coefficient is 35,000 M-1cm-1, quantum yield of 0.54.

HiLyte™ Fluor 488

  • Excellent replacement for FITC
  • Fluorescence of HiLyte™ Fluor 488 is not affected by pH (working pH range is 4-10), a major improvement over fluorescein, which emits its maximum fluorescence only at about pH 9.
  • Protein conjugates prepared with HiLyte™ Fluor 488 dyes are far more superior to conjugates of FITC.
  • HiLyte™ Fluor 488 conjugates are significantly brighter than fluorescein conjugates and are much more photostable.
  • Ex/Em = 497/525 nm, extinction coefficient is 70,000 M-1cm-1, quantum yield of 0.91.

HiLyte™ Fluor 532

  • Excellent replacement for Alexa Fluor™ 532 and Atto 532
  • HiLyte™ Fluor 532 dyes are a class of cyanine dyes.
  • Unlike traditional cyanine dyes, HiLyte Fluor™ 532 dyes are not only highly water-soluble and pH-insensitive but also photostable.
  • HiLyte™ Fluor 532 is suited for the frequency-doubled Nd:YAG laser line (Flow cytometry laser line: 532 nm; Microscopy laser line: 532 nm).
  • Ex/Em = 545/565 nm, extinction coefficient is 170,000 M-1cm-1, quantum yield of 0.26.

HiLyte™ Fluor 555

  • HiLyte™ Fluor 555 conjugates are more photostable and brighter than conjugates of Cy3™, the preferred dye for preparing orange-fluorescent bioconjugates.
  • Compared to the spectra of Cy3™ dyes, the spectra of HiLyte™ Fluor 555 conjugates are slightly red-shifted, resulting in an optimal match of filters designed for Cy3™ dyes.
  • Photostability of HiLyte™ Fluor 555 provides additional time for image capture.
  • Ex/Em = 550/566 nm, extinction coefficient is 150,000 M-1cm-1, quantum yield of 0.10.

HiLyte™ Fluor 647

  • Excellent replacement for Cy5™ and Alexa Fluor™ 647
  • Spectra of HiLyte™ Fluor 647 conjugates are only slightly red-shifted compared to Cy5™, thus HiLyte™ Fluor 647 dye optimal matches filters designed for Cy5™ dyes.
  • In a side-by-side comparison of antibody conjugates of HiLyte™ Fluor 647 dyes and Cy5™ conjugates (supplied by other companies), the total fluorescence of HiLyte™ Fluor 647 labeled secondary antibodies is significantly higher than that of Cy5™ conjugates.
  • HiLyte™ Fluor 647 dyes show only a very little change in absorption or fluorescence spectra when conjugated to most proteins, oligonucleotides, and nucleic acids, thus - unlike Cy5™ dyes - yielding greater total fluorescence at the same degree of substitution.
  • Ex/Em = 649/672 nm, extinction coefficient is 250,000 M-1cm-1, quantum yield of 0.33.

HiLyte™ Fluor 750

  • Excellent replacement for Cy7™
  • Spectrum similar to Cy7™ dye
  • The fluorescence emission maximum of HiLyte™ Fluor 750 is 778 nm which is well separated from commonly used far-red fluorophores, including HiLyte™ Fluor 647, HiLyte™ Fluor 680, or allophycocyanin (APC), facilitating multicolor analysis.
  • With a peak excitation at ~753 nm, conjugates of HiLyte™ Fluor 750 dyes can be excited by a xenon-arc lamp or dye-pumped lasers operating in the 720–750 nm range.
  • Due to its infrared spectra HiLyte™ Fluor 750 is the ideal candidate for in vitro and in vivo tissue imaging.
  • Ex/Em = 750/782 nm, extinction coefficient is 275,000 M-1cm-1, quantum yield of 0.12.
  • Trademarks: HiLyte™ Fluor (AnaSpec), Alexa Fluor™ (Invitrogen), Cy™ (GE Healthcare), DyLight™ (Pierce)


  • 2-Photon/confocal microscopy
  • Flow cytometry
  • In-vivo imaging


  • For shipping and storage information please click on Order#.
Order# Description Amount Price Data Sheet
89317-1AS HiLyte™ Fluor 405 acid, SE 1 mg 149,00 PDF
89319AS HiLyte™ Fluor 405 hydrazide 1 mg 178,00 PDF
81160AS HiLyte™ Fluor 488 acid 10 mg 714,00 PDF
81161AS HiLyte™ Fluor 488 acid, SE 5 mg 429,00 PDF
81162AS HiLyte™ Fluor 488 amine, TFA Salt 1 mg 178,00 PDF
81164AS HiLyte™ Fluor 488 C2 maleimide 1 mg 178,00 PDF
81163AS HiLyte™ Fluor 488 hydrazide 1 mg 178,00 PDF
64348AS HiLyte™ Fluor 488 hydroxylamine, HCl salt *single isomer* 1 mg 338,00 PDF
89340AS HiLyte™ Fluor 532 acid 5 mg 782,00 PDF
89341AS HiLyte™ Fluor 532 acid, SE 1 mg 246,00 PDF
89344AS HiLyte™ Fluor 532 amine 1 mg 229,00 PDF
89342AS HiLyte™ Fluor 532 C2 maleimide 1 mg 229,00 PDF
89343AS HiLyte™ Fluor 532 hydrazide 1 mg 229,00 PDF
81250AS HiLyte™ Fluor 555 acid 5 mg 782,00 PDF
81251AS HiLyte™ Fluor 555 acid, SE 1 mg 246,00 PDF
81252AS HiLyte™ Fluor 555 amine 1 mg 217,00 PDF
81254AS HiLyte™ Fluor 555 C2 maleimide 1 mg 217,00 PDF
81271AS HiLyte™ Fluor 594 acid 10 mg 714,00 PDF
81272-1AS HiLyte™ Fluor 594 acid, SE 1 mg 149,00 PDF
81272-5AS HiLyte™ Fluor 594 acid, SE 5 mg 429,00 PDF
81274AS HiLyte™ Fluor 594 hydrazide - TFA Salt 1 mg 178,00 PDF
81255AS HiLyte™ Fluor 647 acid 5 mg 782,00 PDF
81256AS HiLyte™ Fluor 647 acid, SE 1 mg 246,00 PDF
81257AS HiLyte™ Fluor 647 amine 1 mg 217,00 PDF
81259AS HiLyte™ Fluor 647 C2 maleimide 1 mg 229,00 PDF
81266AS HiLyte™ Fluor 750 acid, SE 1 mg 246,00 PDF
81267AS HiLyte™ Fluor 750 amine 1 mg 229,00 PDF
81269AS HiLyte™ Fluor 750 C2 maleimide 1 mg 229,00 PDF
81268AS HiLyte™ Fluor 750 hydrazide 1 mg 229,00 PDF
  • All prices are in EURO excl. VAT and shipping. Only available in selected countries.