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.

Data and images for MACSPlex Cytokine Kits

Figures

Figure 1: Principle of MACSPlex Cytokine 12 Kit, human and MACSPlex Cytokine 10 Kit, mouse

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Figure 2: Principle of MACSPlex Cytotoxic T/NK Cell Kit, human

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Legend Figure 1 + 2.
Samples containing unknown levels of analytes are incubated with antibody-coated MACSPlex Beads (MPx), and the analytes (e.g., Cytokines) bind to the specific antibodies. The analytes are labeled with APC-conjugated antibodies, the Detection Reagent, either directly (Figure 1) or indirectly via biotinylated antibodies (Figure 2)
Consequently, sandwich complexes are formed between the MACSPlex Bead, the Cytokine and the Detection Reagent. These complexes can be analyzed based on the fluorescence characteristics of both the MACSPlex 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 the samples.
Legend Figure 1 + 2.
Samples containing unknown levels of analytes are incubated with antibody-coated MACSPlex Beads (MPx), and the analytes (e.g., Cytokines) bind to the specific antibodies. The analytes are labeled with APC-conjugated antibodies, the Detection Reagent, either directly (Figure 1) or indirectly via biotinylated antibodies (Figure 2)
Consequently, sandwich complexes are formed between the MACSPlex Bead, the Cytokine and the Detection Reagent. These complexes can be analyzed based on the fluorescence characteristics of both the MACSPlex 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 the samples.

Specifications for 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.

Detailed product information

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.

References for MACSPlex Cytokine Kits

Publications

  1. Yang, Y. et al. (2020) Human umbilical cord mesenchymal stem cells ameliorate skin fibrosis development in a mouse model of bleomycin-induced systemic sclerosis. Exp Ther Med. 20(6): 257
  2. Piermattei, A. et al. (2016) Toll-Like Receptor 2 Mediates In Vivo Pro- and Anti-inflammatory Effects of Mycobacterium Tuberculosis and Modulates Autoimmune Encephalomyelitis. Front Immunol 7: 191
  3. Tran, H.T.T. et al. (2021) Human T2R38 Bitter Taste Receptor Expression in Resting and Activated Lymphocytes Front Immunol 12: 669005
  4. Jureczek, J. et al. (2020)
    Highly Efficient Targeting of EGFR-Expressing Tumor Cells with UniCAR T Cells via Target Modules Based on Cetuximab
    ®
    Onco Targets Ther. 13: 5527
  5. Bachmann, D. et al. (2017) Retargeting of UniCAR T cells with an in vivo synthesized target module directed against CD19 positive tumor cells. Oncotarget. 9(7): 7487-7500
  6. Foltz, J. A. et al. (2018) TGFβ Imprinting During Activation Promotes Natural Killer Cell Cytokine Hypersecretion Cancers (Basel) 10(11): 423
  7. Barone, M. et al. (2021) The role of circulating monocytes and JAK inhibition in the infectious-driven inflammatory response of myelofibrosis Oncoimmunology 9(1): 1782575
  8. Loureiro, L.R. et al. (2018) Development of a novel target module redirecting UniCAR T cells to Sialyl Tn-expressing tumor cells Blood Cancer J. 8(9): 81
  9. Leong, T. et al. (2020) Preclinical Activity of Embryonic Annexin A2-Specific Chimeric Antigen Receptor T Cells Against Ovarian Cancer Int J Mol Sci 21(2): 381
  10. Pfeiffer, A. et al. (2018) In vivo generation of human CD19-CAR T cells results in B-cell depletion and signs of cytokine release syndrome EMBO Mol. Med. 10(11)
  11. Xiang, Y et al. (2019) In vitro expansion affects the response of human bone marrow stromal cells to irradiation. Stem Cell Res Ther. 10(1): 82
  12. Rosshirt, N. et al. (2021) Proinflammatory T cell polarization is already present in patients with early knee osteoarthritis Arthritis Res Ther. 23(1): 37
  13. Lantier, L. et al. (2020) Neospora caninum: a new class of biopharmaceuticals in the therapeutic arsenal against cancer. J Immunother Cancer 8(2): e001242
  14. Conde, E. et al. (2021) Dual vaccination against IL-4 and IL-13 protects against chronic allergic asthma in mice Nat Commun. 12(1): 2574
  15. Eliasse, Y. et al. (2019) Effect of thermal spring water on human dendritic cell inflammatory response. J Inflamm Res. 12: 181

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