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CF™532 Dye

CF™532 is an orange colored fluorescent dye with absorption and emission maxima at 527 and 558 nm, respectively (Figure 1). The dye is based on a new class of rhodamine dyes invented by Biotium scientists. Unlike traditional rhodamine dyes, CF™532 is not only bright and photostable but also highly water-soluble. As a result, CF™532 antibody conjugates retain excellent solubility and yield the brightest fluorescence among antibody conjugates prepared from spectrally similar dyes. For example, CF™532 labeled goat anti-mouse IgG is significantly brighter than the same antibody labeled with Alexa Fluor® 532 at a similar degree of labeling (Figures 2, 3).
A full selection of reactive dyes, secondary antibodies, antibody labeling kits, and other bioconjugates including phalloidins, Annexin V and α-bungarotoxin are available for CF™ dyes.

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CF™532 conjugates, labeling kits, and reactive dyes

 

Technical Summary

  • Abs/Em Maxima: 527/558 nm
  • Extinction coefficient: 96,000
  • Molecular weight: ~685
  • Flow cytometry laser line: 532 nm
  • Microscopy laser line: 532 nm
  • Direct replacement for: Alexa Fluor® 532, Atto 532

Advantages

  • Significantly brighter than Alexa Fluor® 532
  • Highly water-soluble and pH-insensitive
Figure 1. Absorption and emission spectra of CF™532 conjugated to goat anti-mouse IgG in PBS.
Figure 1. Absorption and emission spectra of CF™532 conjugated to goat anti-mouse IgG in PBS.

Figure 2. Comparison of relative fluorescence of goat anti-mouse IgG antibody labeled with CF532 or Alexa Fluor® 532 at various degrees of labeling (DOL) (number of dye molecules per antibody).
Figure 2. Comparison of relative fluorescence of goat anti-mouse IgG antibody labeled with CF532 or Alexa Fluor® 532 at various degrees of labeling (DOL) (number of dye molecules per antibody).
Figure 3. Flow cytometry analysis of Jurkat cells stained with Alexa Fluor® 532 antibody or CF™532 secondary antibody conjugates. Intracellular staining was performed with mouse anti-CD3 antibody followed by goat anti-mouse IgG conjugates. Background was determined by staining with secondary antibody (2nd Ab) alone. Fluorescence was analyzed on a BD FACSCalibur flow cytometer in the FL2 channel. The bars represent the relative fluorescence of the geometric means of the population of cells.
Figure 3. Flow cytometry analysis of Jurkat cells stained with Alexa Fluor® 532 antibody or CF™532 secondary antibody conjugates. Intracellular staining was performed with mouse anti-CD3 antibody followed by goat anti-mouse IgG conjugates. Background was determined by staining with secondary antibody (2nd Ab) alone. Fluorescence was analyzed on a BD FACSCalibur flow cytometer in the FL2 channel. The bars represent the relative fluorescence of the geometric means of the population of cells.