Supernatants from whole blood stimulated for 18 hours were thawed. Concentrations of IL-12p40, IL-12p70, IL-10, IL-6 and TNF-α were measured for all samples by Milliplex MAP Multiplex Immunoassay (based on Luminex MAP technology; Millipore Corporation) on a Bio-Rad Luminex 100 Bio-Plex Liquid Array Multiplexing System Fluorescent Reader, according to the manufacturer’s instruction. Concentrations of the standards in 200µL assay buffer ranged from 10000, 2000, 400, 80, 16 to 3.2 pg/mL. A sample volume of 25µL either diluted 2- or 25-fold was used.
Luminex 100 bio plex liquid array multiplexing system fluorescent reader
Manufactured by Bio-Rad
The Luminex 100 Bio-Plex Liquid Array Multiplexing System Fluorescent Reader is a laboratory instrument designed for the detection and quantification of multiple analytes in a single sample. The system utilizes fluorescent-coded beads to simultaneously measure a variety of biomolecules, such as proteins, cytokines, and nucleic acids.
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3 protocols using luminex 100 bio plex liquid array multiplexing system fluorescent reader
1
Multiplex Cytokine Quantification
2
Multiplex Cytokine Analysis of BCG-Stimulated Blood
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Supernatants from whole blood stimulated for 6 days with BCG were thawed. Concentrations of IL-1β, IL-4, IL-6, IL-10, IL-17, IL-22, IL-23, IFNγ, and TNFα were measured for all samples by Milliplex MAP Multiplex Immunoassay (based on Luminex MAP technology; Millipore) on a Bio-Rad Luminex 100 Bio-Plex Liquid Array Multiplexing System Fluorescent Reader, according to the manufacturer’s instruction. The following concentrations of the standards in 200 ml assay buffer were used: 10,000, 2,000, 400, 80, 16, and 3.2 pg/ml.
3
Multiplex Cytokine Profiling in Whole Blood
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Concentrations of IL1β, IL4, IL6, IL10, IL17, IL22, IL23, IFNγ, TNFα were measured in supernatants from whole blood stimulated for 6 days by Milliplex MAP Multiplex Immunoassay (based on Luminex MAP technology; Millipore) on a Bio-Rad Luminex 100 Bio-Plex Liquid Array Multiplexing System Fluorescent Reader, according to the manufacturer’s instructions. The following concentrations of the standards diluted in 200 μl assay buffer were used: 10000, 2000, 400, 80, 16, and 3.2 pg/ml.
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