Jun 13, 2023
Luminex-Based Multiplex Cytokine Analysis
- michela.deleidi 1,2,
- Bianca Marchetti3,4,
- Federico Bertoli1,2,
- Carmela Giachino3
- 1Mitochondria and Inflammation in Neurodegenerative Diseases, DZNE, Tübingen-Germany;
- 2Hertie Institute for Clinical Brain Research, University of Tübingen;
- 3Neuropharmacology Laboratory, Oasi Research Institute-IRCCS, Troina, Italy;
- 4Biomedical and Biotechnological Sciences, Pharmacology Section, University of Catania-Italy
Protocol Citation: michela.deleidi , Bianca Marchetti, Federico Bertoli, Carmela Giachino 2023. Luminex-Based Multiplex Cytokine Analysis. protocols.io https://dx.doi.org/10.17504/protocols.io.ewov1ob7plr2/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: March 31, 2023
Last Modified: June 13, 2023
Protocol Integer ID: 79821
Keywords: Luminex-Based Multiplex Cytokine Analysis, based multiplex cytokine analysis multiplexed kit, multiplex cytokine analysis multiplexed kit, based multiplex assay, chemokine biomarker with the high sensitivity bead, multiplex assay, luminex technology, chemokine biomarker, mouse serum, luminex, high sensitivity bead, cell culture sample
Funders Acknowledgements:
ASAP-Aligning Science Across Parkinson’s
Grant ID: ASAP-000420
Abstract
Multiplexed kit "Mouse High Sensitivity T Cell Magnetic Bead Panel" allows a Simultaneous analyze low levels of cytokine and chemokine biomarker with the High Sensitivity Bead-Based Multiplex Assays using the Luminex technology, in mouse serum, plasma and cell culture samples.
Attachments
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Materials
Materials
- Mouse High Sensitivity T Cell Magnetic Bead Panel" (MHSTCMAG-70K, MILLIPLEX, EMD Millipore Corp., Billerica, MA, USA)
- FLEXMAP 3D® instrument (Luminex Corp., Austin, TX, USA)
Luminex-Based Multiplex Cytokine Analysis
Measure Cytokines from murine plasma samples, including IFN-γ, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-17A, and TNF-α simultaneously using the multiplexed kit "Mouse High Sensitivity T Cell Magnetic Bead Panel" (MHSTCMAG-70K, MILLIPLEX, EMD Millipore Corp., Billerica, MA, USA).
Perform the assay according to the manufacturer's instructions with an Overnight incubation step (17 hours) at 4 °C .
Analyze samples as single values and use two controls with corresponding acceptance ranges, provided with the kit, for quality control purposes.
Perform the readout on a FLEXMAP 3D® instrument (Luminex Corp., Austin, TX, USA).
Acquire data using Luminex xPONENT® software (version 4.3) and determine mean fluorescence intensity (MFI).
Use the Bio-Plex ManagerTM software (version 6.2) (Bio-Rad, Hercules, CA, USA) for back-calculation of unknown cytokine concentrations.