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The Luminex 100 reader is a flexible platform based on flow cytometry that enables multiplex analysis of up to 100 analytes in a single microplate well. The system is capable of analyzing various biomarkers including nucleic acids, receptor-ligand assays, immunoassays and enzymatic assays. The platform is based on the simultaneous detection of individual fluorescently dyed microspheres and antibodies conjugated to phycoerythrin (PE). In our lab, we use the Luminex 100 reader in several different assays including for HLA class I typing, KIR typing and antibody specificity determination. The assay described below uses the Luminex 100 reader to determine the antibody specificity of a monoclonal anti-HLA class I antibody.
Take care not to allow the sheath fluid resevoir to run dry as allowing air into the FACS system can lead to aberrant results and damage to the instrument. Sheath fluid may need re-filling if greater than 48 samples are being analyzed in a single experiment. Calibration should be undertaken prior to each use of the instrument to ensure the correct identity of all 96 bead types.