BioLegend LEGENDplex
BioLegend’s LEGENDplex™ bead-based immunoassays quantify multiple soluble analytes simultaneously in biological samples using a flow cytometer.
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BioLegend's LEGENDplex™ kits are provided as predefined panels, ranging from 3 to 13 specificities, or customers can mix and match any subset within each predefined panel using our Mix and Match system. If you would like to combine specificities across different predefined panels, i.e., Human Cytokine and Human Chemokine, please visit our custom panels tab at the top of this page.

Each kit provides sufficient reagents for at least 100 tests. The assay can be performed in a 96-well filter plate, allowing you to run 40 samples plus 8 standard curve titrations in duplicate, or they can be performed in FACS tubes, where duplicate titrations may not be necessary.

For analysis of LEGENDplex™ assays, BioLegend provides both PC and Mac versions of the LEGENDplex™ data analysis software free of charge. To download the software, click on the Software tab above. Note: full use of software functions requires a USB dongle (included in each kit) for PC users, or a no cost license key (see installation guide) for Mac users.
Principle of the Assay
BioLegend's LEGENDplex™ is a bead-based immunoassay that utilizes the same basic principles of sandwich immunoassays, whereby a soluble analyte is captured between two antibodies. 
  1. Bead populations come in two sizes and differing levels of APC fluorescence, allowing them to be clearly distinguished from each other. Each bead set is conjugated with a specific antibody on the surface and serves as the capture bead for that particular analyte. When a selected panel of capture beads are mixed together and incubated with an unknown sample containing target analytes, each analyte will be bound by its specific capture bead.
  2. After washing, biotinylated detection antibodies are added and each detection antibody will bind to its specific analyte bound on the capture beads, thus forming capture bead-analyte-detection antibody sandwiches.  Streptavidin-phycoerythrin (SA-PE) is subsequently added, which will bind to the biotinylated detection antibodies, providing fluorescent signal with intensities in proportion to the amount of bound analyte.
  3. For each bead population, the PE signal fluorescence intensity is then quantified using a flow cytometer.
  4. The concentration of a particular analyte is determined based on a known standard curve using the LEGENDplex™ data analysis software.
  • Compatible with standard flow cytometers
  • Free data analysis software
  • Excellent assay performance
  • Outstanding pricing
View a detailed LEGENDplex™
assay protocol video on JoVE.

Currently available reactivitiesHuman, Mouse, Rat, Non-Human Primates
Maximum number of analytes in a plex13
Minimum required sample volume12.5 µl serum/plasma per well, 25 µl for tissue culture samples
Instrument requiredFlow cytometer capable of detecting PE and APC (or PerCP/FL3 on instruments without a red laser)
Tests per kit100 tests
Want to get hands-on training on how to perform the assays and analyze your data using LEGENDplex™ products?

Request a free demo at your lab with one of our Field Application Scientists or come visit us at BioLegend headquarters in San Diego!

Laboratory Demo Details:

  • One of our Field Application Scientists will visit your lab to assist you with hands-on product usage at the bench, including setup and sample acquisition on flow cytometers, as well as data analysis using LEGENDplex™ software.
  • Run a LEGENDplex™ assay using your own biological samples.
  • Assay demonstrations are provided free of charge to the laboratory.

BioLegend HQ Visit Details:

  • One or two days of training, depending on your schedule, at BioLegend headquarters in sunny San Diego.
  • Hands on product usage at the bench, including setup and sample acquisition on flow cytometers, as well as data analysis using LEGENDplex™ software.
  • Plenty of time for troubleshooting and discussions with product development scientists.
  • Bringing your own samples is highly encouraged.
  • There is no charge for the training.
  • Housing and meals are not provided. Customers are responsible for their own travel and housing accommodations.
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