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Near-Infrared CF™ Dyes

Comparison of microtubule imaging using conventional wide-field microscopy (left) with STORM (right) using CF™ dye conjugates. Images courtesy of Sam Kenny and Professor Ke Xu, College of Chemistry, University of California, Berkeley.

Next generation near-infrared dyes

Near-infrared (near-IR) dyes offer important advantages over traditional visible light dyes. Because cellular or tissue components produce minimal autofluorescence in the near-IR region, near-IR dyes have the potential to offer highly specific and sensitive detection in complex biological systems. Light with wavelength in the near-IR region has strong tissue penetration, allowing the use of near-IR dyes for in vivo fluorescence imaging. For Western blotting, near-IR dyes offer advantages over chemiluminescence in sensitivity, linearity, and multi-color detection.

Near-IR CF™ dyes are next-generation long wavelength dyes representing a true breakthrough in the field. The current commercial near-IR dyes suffer from problems of limited fluorescence brightness due to excessive dye aggregation and poor stability. As a result of novel molecular engineering by scientists at Biotium, near-IR CF™ dyes overcome these problems, resulting in exceptionally bright and photostable dyes. In addition, near-IR CF™ dyes possess a proprietary structural feature that renders the dyes more water-soluble than other near-infrared dyes without introducing excessive charge. Consequently, a higher number of near-IR CF™ dyes can be conjugated to a protein for maximal fluorescence, while still preserving conjugate specificity and long in vivo half-life.

Near-IR CF Dyes

Biotium currently offers 6 NIR CF dyes, named by their peak excitation wavelengths. Our NIR CF dyes have industry-leading brightness and low background.

  • CF™680
  • CF™680R
  • CF™750
  • CF™770
  • CF™790
  • CF™800

Advantages

  • Brightness: Near-IR CF™ dyes match or surpass the brightness of Alexa Fluor®, Cy® dyes and others
  • Photostability: Near-IR CF™ dyes are among the most photostable dyes
  • pH Insensitivity: CF™ dyes remain highly fluorescent over a wide pH range
  • Solubility: CF™ dyes are highly soluble in both water and a wide range of organic solvents
  • Specificity and In Vivo Stability: CF™ dye conjugates offer super specificity and improved half-life for in vivo imaging
  • Compatibility: Compatible with LI-COR Odyssey® and other near-IR imaging systems

Applications

  • In Vivo Imaging: Reactive dyes and kits for small animal imaging
  • Protein and Antibody Labeling: Reactive dyes and kits for custom conjugations
  • Westerns: Perform two-color Westerns with a wide-selection of highly cross-adsorbed secondary antibodies conjugated to CF™680 and CF™770
  • Microscopy: Bright and photostable antibodies or reactive dyes for multicolor imaging
  • Super-Resolution Microscopy: CF™680 and CF™750 have been validated in STORM, CF680R is compatible with STED – learn more about CF™ Dyes for Super-Resolution Imaging

View products associated with this technology…

Near-IR CF™ dye conjugates, labeling kits, and reactive dyes

 

NIR CF dyes spectra
Absorption and emission spectra of near-infrared CF dyes.

 


CF™ DyeEx/Em (nm)Replacement forAdvantagesView Products
CF™680681/698Alexa Fluor® 680, Cy®5.5, DyLight® 680, IRDye® 680LT, IRDye® 680RD• Brightest 680 nm dye
• Superior signal-to-noise ratio for immunostaining
CF™680 Products
CF™680R680/701Alexa Fluor® 680, Cy®5.5, DyLight® 680, IRDye® 680LT, IRDye® 680RD• Most photostable 680 nm dye
• Suitable for labeling nucleic acids and small biomolecules
CF™680R Products
CF™750755/777Alexa Fluor® 750, Cy®7,
DyLight® 750, APC-Alexa Fluor® 750, IRDye® 750
• Exceptionally bright and stable
• Highly water soluble
• Non-tandem alternative to APC-Alexa Fluor® 750
CF™750 Products
CF™770770/797DyLight® 800, IRDye® 800CW• Exceptionally bright and stable
• Highly water soluble
CF™770 Products
CF™790784/806Alexa Fluor® 790• Exceptionally bright and stable
• Highly water soluble
CF™790 Products

Applications: Western Blotting

IR680LT IR800CW vs CF680 CF770 blot labeled
Western detection with near-IR secondary antibody conjugates. HeLa cell lysate (2 ug-0.125 ug total protein, lanes 1-5) was probed with mouse anti-tubulin and rabbit anti-COXIV, followed by goat anti-mouse (red) and goat anti-rabbit (green) conjugated to IRDye 680LT and IRDye 800CW (left) or CF680 and CF770 (right) at the same final concentrations. Membranes were scanned using an Odyssey infrared imaging system (LI-COR Biosciences). CF dyes were 1.5-2 fold brighter than IR dyes by band quantitation.

 

Advantages: Brightness

CF770 conjugation
Relative fluorescence of goat anti-mouse IgG conjugated to CF770, DyLight 800, or IRDye 800 CW at varying degrees of labeling (dye molecules per antibody molecule).

Applications: In Vivo Imaging

CF750 in vivo
Tumors in mice were imaged using an IVIS system (Caliper Life Sciences) 24-96 hours after injection of Avastin antibody conjugated to CF750. Images courtesy of Caliper Life Sciences.

 

Advantages: Specificity

CF790 Western specificity
Near-IR CF dyes carry less negative charge than other near-IR dyes, resulting in less non-specific binding of conjugates. Western blots of HeLa cell lysate (lane 2) probed with mouse anti-tubulin followed by goat anti-mouse conjugated to Alexa Fluor 790 (left) or CF790 (right). Lane 1: near-IR protein marker (LI-COR Biosciences).

*CF dye technology is covered by pending US and international patents; Alexa Fluor is a registered trademark of Invitrogen; Cy Dye is a trademark of GE Healthcare; DyLight is a trademark of Thermo Fisher Scientific; IRDye and Odyssey are registered trademarks of LI-COR Biosciences; In-Cell Western is a trademark of LI-COR Biosciences.