Multiplex Assays

At Green Mountain Antibodies, we feel it is important to understand the specificity of your antibody.  We use a multiplex flow cytometry technology which allows for detection of up to 100 proteins in one assay!  

Detection of antibodies using multiplex technology relies on a unique combination of conditions.  The first of these is the microsphere, or bead.  Different antigens can be coupled to over 100 different sets of beads.  One set can be distinguished from another because each contains a characteristic ratio of two dyes.  All beads that have a particular color ratio belong to a bead set, which is identified by a bead ID number.  One can then couple each bead set to a particular protein.  In order for a bead to be detected by the instrument, it must first be coupled to an antigen, which then must bind to an antibody in the sample plasma, which then must bind to a secondary fluorochrome-conjugated antibody.  If these four elements are not present, the bead will not be read by the lasers.  Because there are 100 different color ratios, each with their own location on the bead map, up to 100 bead sets can be mixed together allowing the multiplex instrument to sort the data.  The amount of a particular antibody present in a sample is displayed as the median fluorescence intensity. This value is a result of the instrument averaging the intensity of beads 50 and 51 out of 100 beads from each set. 

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