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Fluorophore Guide

Antibodies Conjugated to Fluorophores

Bio-Rad offers a wide range of antibodies conjugated to all of the most commonly used fluorophores.

In addition to FITC, PE, and APC, we also offer Alexa Fluor® and DyLight® conjugates in an array of colors, both with outstanding spectral properties. You can use our Search to see if we have the antibody you are working with, labeled to suit your needs.

If you can’t find the format you need, just let us know. If we don’t have the labeled version you are seeking, we offer easy-to-use LYNX Rapid Conjugation Kits® for many popular dyes and tandem conjugates as well as custom conjugation services.

ffer a wide range of antibodies conjugated to all of the most commonly used fluorophores. In addition to FITC, PE, and APC, we also offer Alexa Fluor® and DyLight® conjugates in an array of colors, both with outstanding spectral properties. You can use our Search to see if we have the antibody you are working with, labeled to suit your needs. If you can’t find the format you need, just let us know. If we don’t have the labeled version you are seeking, we offer easy-to-use LYNX Rapid Conjugation Kits® for many popular dyes and tandem conjugates as well as custom conjugation services.
- See more at Bio-Rad: https://www.bio-rad-antibodies.com/fluorophores.html?utm_source=shared&utm_medium=referral&utm_campaign=page-copy-copied-and-pasted

Single Dyes for Flow Cytometry / Microscopy

Tandem Dyes for Flow Cytometry

When designing panels of eight or more colors, tandem dyes, such as APC-Cy7, have to be included. This is due to both laser excitation and single fluorophores limitations, which make it necessary for a single laser to excite the maximum number of fluorophores possible.

Tandem dyes, as the name implies, consist of two different covalently attached fluorophores (a donor and an acceptor molecule). With regards to spectral properties, the tandem dye has the excitation characteristics of the donor fluorophores and the emission characteristics of the acceptor molecule. These properties are due to Förster resonance energy transfer (FRET; also known as Fluorescence resonance energy transfer); a process in which energy is passed on from an excited donor to a nearby acceptor molecule, which then emits a photon of light.

For tandem dyes the following guidelines should be followed:

Tandem dyes are highly susceptible to photobleaching and therefore need to be protected from light at all times.
Tandem dye antibody conjugates should never be frozen. Freezing might result in denaturing of the donor fluorophores and thereby no or reduced staining.
The brightness of tandem dyes might be reduced by a fixation or permeabilization step. If fixation or permeabilization is required, the duration should be as short as possible.
Tandem dyes have a high batch-to-batch variation and therefore each batch has to be re-optimized. Tandem dyes are highly variable between different suppliers.

Abbreviations
APC	Allophycocyanin
FITC	Fluorescein isothiocyanate
PE	Phycoerythrin (Note. Phycoerythrin (PE) is same as R-Phycoerythrin (RPE)
PerCP	Peridinin-chlorophyll-protein complex

Alexa Fluor® and Pacific Blue™ are trademarks of Molecular Probes Inc, OR, USA.
DyLight® is a trademark of Thermo Fisher Scientic and its subsidiaries.
LYNX Rapid Conjugation Kits® is a registered trademark of Bio-Rad AbD Serotec Ltd.
Cy containing products or portions thereof are manufactured under licence from Carnegie Mellon University under U.S. Patent Number 5,268,486 and related patents.
Cy™ and CyDye™ are trademarks of GE Healthcare Limited
For research purposes only. Not for therapeutic or diagnostic use