Technical Information

Technical Information

Technical Notes: Multi-Cytokine Profiling

Unlike conventional immunoassays, R&D Systems’ Fluorokine® MAP kit can run multiple cytokine immunoassays on a single heterogeneous sample. This multiplexed assay is designed using the Luminex™ LabMAP™ detection technology. Luminex LabMAP latex microbeads are dyed with two fluorophores. Precise ratios of the two fluorophores create distinguishable bead sets. These beads are the solid phase for the Fluorokine MAP multiplex immunoassay.

Cytokine capture antibodies are coupled to specific bead sets. Each unique antibody-coated bead set is then an "individual cytokine immunoassay ". Various antibody-coated bead sets can be mixed together (i.e. multiplexed) and incubated with a heterogeneous sample. After cytokines bind to their corresponding capture antibody/bead, a Phycoerythrin (PE)-conjugated cytokine-specific detection antibody is used as the reporter. The amounts of cytokines bound are proportional to the PE signals generated for each bead set. The fluorescence levels generated by the beads and the PE-labeled antibody are analyzed on a Luminex 100 instrument using a dual laser system as they pass through a flow cell. Real time quantitative data is generated for each signal.

Features of Fluorokine MAP

  • Analytes: IL-1 beta, IL-2, IL-4, IL-6, IL-8, IL-10, GM-CSF, TNF-alpha, FGF basic, G-CSF, IFN-gamma, VEGF
  • Sample Type: Cell culture supernate
  • Sample Size: 50 µL
  • Sensitivity: Generally < 10 pg/mL
Figure 1
Figure 1. Typical standard curves for IL-1 beta, IL-10, and IL-6 run simultaneously.
Analyte Kit Contents
  • Concentrates of cytokine-specific capture antibodies coated on beads and directly conjugated cytokine-spcific detection antibodies
  • Each analyte kit is sold separately or as a multiplexed set (in any combination)
Base Kit Contents
  • Multiplex standard
  • Bead and calibrator diluents
  • Wash buffer
  • 96-well filter-bottomed microplate
  • Plate sealers

Luminex and LabMAP are registered trademarks of Luminex Corp.

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