LEGENDplex™ Multiplex Bead Assays
Current multiplex assays are frequently associated with high reagent and instrument costs and difficulties with performance and data analysis. To address these issues, BioLegend is proud to offer LEGENDplex™ panels for the simultaneous detection of human or mouse cytokines and chemokines.
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.
LEGENDplex™ Data Analysis Software
The software can be used as a trial version with limited functionality for 30 days. A USB dongle enables full use of software functions. Each dongle provides a limited number of analysis runs. A dongle is provided free of charge with each purchase of a LEGENDplex™ product. Now available for Windows and MAC.
Example Data
Panels
