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Biolegend Simple Rules for Polychromatic Assays

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Multicolor immunofluorescent staining is essential in identifying mixed cell populations or characterizing multiple parameters in single cells by flow cytometry or immunofluorescence microscopy. Tandem dyes expand the possibilities for fluorescence color selection of antibodies for use in multicolor flow cytometry.

These tandem dyes can provide much higher wavelength fluorescence emission relative to the excitation wavelength and thus allows for multiple distinct emission ranges from a single laser.

BioLegend is proud to announce our expanding list of products newly conjugated to PerCP and PerCP/Cy5.5!   Click here to view all of our PerCP and PerCP/Cy5.5 products.

BioLegend conveniently provides antibodies with the following tandem dyes: PE-Cy5, PE-Cy5.5, PE-Cy7, PerCP/Cy5.5, APC/Cy5.5, and APC/Cy7 in addition to our standard dyes: FITC, PE, Alexa Fluor® 488, Alexa Fluor® 647, Alexa Fluor® 700, PerCP, APC, and Pacific Blue.  If you don’t find the tandem dye you are looking for attached to your antibody of choice, just request a custom conjugation.

Introducing BioLegend's Spectra Analyzer try it out today!

Multicolor Staining

Choosing antibodies and fluorochromes for multicolor flow cytometry can be confusing and frustrating. Use our Guide Wheel to assist you in developing and optimizing your flow cytometry experiments. Master the five aspects of the Guide Wheel and you will be well on your way to legendary discovery.

  1. Know your instrument and the fluorochromes that you can use.
  2. Learn about the fluorochrome brightness index.
  3. Learn the Rules for choosing and using fluorochromes.
  4. Know other resources you can use.
  5. Learn about the expression of common cell surface proteins.

View our Resource Guides and Protocols

Flow Cytometry Resources
Multicolor Immunofluorescent Staining Resource Guide
Intracellular Cytokine Staining Resource Guide
Cell Surface Immunofluorescence Staining Protocol
Intracellular Cytokine Staining Protocol
FOXP3 Intracellular Staining Protocol
Need help selecting antibodies for your multicolor panel? Email our expert Technical Service staff


Fluorochrome Excitation/Emission Chart

Determine which fluorochromes can be used on your flow cytometer. Download a PDF of our Fluorochromes for Flow Cytometry and Microscopy poster or request the full size poster and start decorating your lab.

Download our Interactive Human and Mouse CD Molecule Charts

Interactive Human CD Molecule chart

Interactive Mouse CD Molecule chart