Luminex Assay Principle

 

Multiplex Luminex Assay Principle Step 1: The sample is added to color-coded beads, pre-coated with analyte-specific capture antibodies. Biotinylated detection antibodies are added and form an antibody-antigen sandwich.
The sample is added to a mixture of color-coded beads, pre-coated with analyte-specific capture antibodies. The antibodies bind to the analytes of interest. Biotinylated detection antibodies specific to the analytes of interest are added and form an antibody-antigen sandwich. Phycoerythrin (PE)-conjugated streptavidin is added. It binds to the biotinylated detection antibodies.

 

Beads are read on a dual-laser flow-based detection instrument.

Beads are read on a dual-laser flow-based detection instrument, such as the Luminex 200 or FlexMap® analyzer. One laser classifies the bead and determines the analyte that is being detected. The second laser determines the magnitude of the PE-derived signal, which is in direct proportion to the amount of analyte bound.

In addition to flow-based analyzers, magnetic beads can be read using the Luminex MAGPIX® Analyzer. A magnet in the MAGPIX analyzer captures and holds the magnetic beads in a monolayer, while two spectrally distinct light-emitting diodes (LEDs) illuminate the beads. One LED identifies the analyte that is being detected and, the second LED determines the magnitude of the PE-derived signal. Each well is imaged with a CCD camera.

 

 

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