Background information
MACSPlex Cytokine 12 Kit, human, MACSPlex Cytotoxic T/NK Cell Kit, human and MACSPlex Cytokine 10 Kit, mouse include fluorescently labeled MACSPlex Capture Bead populations that have been coated with antibodies reacting with one of the respective analytes within the sample. The bead populations can be distinguished by different fluorescence intensities detected in the FITC (B1) and PE (B2) channel of the MACSQuant® Analyzer, MACSQuant Analyzer 10 and MACSQuant Analyzer 16 (as well as respective channels of comparable flow cytometers). MACSPlex Capture Beads are added to the samples and to serial dilutions of the MACSPlex Cytokine Standard.
During a 2-hour incubation period, the cytokines are captured by the MACSPlex Capture Beads. MACSPlex Cytokine Detection Reagents, composed of a cocktail of APC-conjugated antibodies recognizing the different analytes, are added in order to form sandwich complexes during a 1-hour incubation period using the MACSPlex Cytokine 12 Kit, human and MACSPlex Cytokine 10 Kit, mouse. With the MACSPlex Cytotoxic T/NK Cell Kit, human APC labeling of the analytes is performed indirectly using analyte-specific biotinylated antibodies and APC-conjugated Biotin antibodies during a total incubation time of 1,5-hours.
Consequently, sandwich complexes are formed between the MACSPlex Capture Bead, the analyte and the detection reagent. These complexes can be analyzed based on the fluorescence characteristics of both the MACSPlex Capture Bead and the detection reagent. Standards of known quantities of given analytes are provided with the Kit and are used for the quantification of the unknown concentrations of analytes within samples. Standard curves for each of the cytokines are generated. The median of the APC fluorescence of each capture bead population gives the concentration of each cytokine in the sample.
Applications
The MACSPlex Cytotoxic T/NK Cell Kit, human has been developed for the simultaneous flow cytometric detection of soluble GM-CSF, Granzyme B, IFN-γ, IL-2, IL-4, IL-6, IL-10, IL-17A, IL-21, MCP-1, Perforin, and TNF-α in a single sample. The kit has been optimized for use with serum, plasma, and cell culture supernatant.
The MACSPlex Cytokine 12 Kit, human has been developed for the simultaneous flow cytometric detection of the soluble human cytokines GM-CSF, IFN-α, IFN-γ, IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12p70, IL-17A, and TNF-α in a single sample. The kit has been optimized for use with serum, plasma, and cell culture supernatants.
MACSPlex Cytokine 10 Kit, mouse has been developed for the simultaneous flow cytometric detection of the soluble mouse cytokines GM-CSF, IFN-γ, IL-2, IL-4, IL-5, IL-10, IL-12p70, IL-17A, IL-23, and TNF-α in a single sample. The kit has been optimized for use with serum, plasma, and cell culture supernatants.
Overview of available MACSPlex Cytokine Kits
Overview
MACSPlex Assays are designed for determining concentrations of multiple soluble analytes in a single sample. The analysis is based on MACSPlex Capture Beads, which display defined fluorescence properties and can be identified using standard flow cytometry techniques. MACSPlex Kits contain a cocktail of various MACSPlex Capture Beads populations, each coated with a specific antibody reacting with one of the respective analytes within the sample. Analytes are labeled directly or indirectly (via biotin) with APC-conjugated antibodies, depending on the chosen Kit. Standards of known quantities of given analytes are provided with the kit and are used for the quantification of the unknown analyte concentrations within the samples.
In combination with the Express Modes of the MACSQuant Analyzers, the MACSPlex Cytokine Kits are optimized for automated measurement of beads. They simplify flow cytometric analysis via predefined experiment settings as well as acquisition and analysis templates. They apply an automated gating strategy to populations of interest that will be automatically adjusted for each data file individually to achieve optimal results.
Besides the MACSQuant Analyzers, the MACSPlex Cytokine Kits are compatible with every other flow cytometer equipped with a blue (488 nm) and red (635 nm) laser, able to discriminate FITC, PE, and APC fluorescence channels. For automated quantitative analysis of MACSPlex data independent of MACSQuant Analyzer Express Modes, the MACSPlex InspectoR Tool is offered. The MACSPlex InspectoR is a free to use online tool providing a comprehensive automated analysis of complex multiplex data. MQD or FCS files can be uploaded, analyzed, and quantitative results downloaded: MACSPlex InspectoR tool | Miltenyi Biotec | Deutschland
