Luminex 200 system

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Luminex 200 system is a versatile multiplex assay platform that can simultaneously analyze multiple analytes in a single sample. The system utilizes color-coded magnetic microspheres, flow cytometry, and dedicated software to enable rapid and accurate quantification of a wide range of biomolecules.

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41 protocols using luminex 200 system

Multiplex Cytokine Quantification in BAL

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Measurement of cytokines in the BAL was performed taking advantage of a well-established method, Luminex® xMAP technology, that allows measuring a panel of multiple analytes on a small volume sample (100 μL) simultaneously [30 (link)]. The assay, for the quantitative detection of IL-4, IL-5, IL-10, and IL-13, was performed using a Milliplex Cytokine Panel plate (Merck Millipore) according to the manufacturer's instructions on automated immunoassay analyzer (Luminex 200™ System, Thermo Fisher Scientific) as previously described [31 (link)]. All samples were run in duplicate. After the run, data were analyzed by using Xponent software (1.9 version, Luminex 200 System; Thermo Fisher Scientific) and the final concentration of each cytokine was expressed in pg/mL.

Spaziano G., Cappetta D., Urbanek K., Piegari E., Esposito G., Matteis M., Sgambato M., Tartaglione G., Russo R., De Palma R., Rossi F., De Angelis A, & D'Agostino B. (2016). New Role of Adult Lung c-kit+ Cells in a Mouse Model of Airway Hyperresponsiveness. Mediators of Inflammation, 2016, 3917471.

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Multiplex Cytokine Analysis in BAL

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Measurement of cytokines in the BAL were performed taking advantage of a well-established method, Luminex xMAP technology (Luminex^® 200^™ System, Life technologies), that allows to measure a panel of multiple analytes on a small volume sample (100 μl) simultaneously [45 (link)]. The assays, for the quantitative detection of IL-4, IL-5, IL-10 and IL-13, were performed using a Milliplex Cytokine Panel plate (Millipore-Merck, Italy) according to the manufacturer’s instructions on automated immunoassay analyzer as previously described [46 (link)]. All samples were run in duplicate. After the run, data were analyzed using by Xponent software (1.9 version, Luminex^® 200^™ System, Life technologies) and the final concentration of each cytokine expressed in pg/ml.

Urbanek K., De Angelis A., Spaziano G., Piegari E., Matteis M., Cappetta D., Esposito G., Russo R., Tartaglione G., De Palma R., Rossi F, & D’Agostino B. (2016). Intratracheal Administration of Mesenchymal Stem Cells Modulates Tachykinin System, Suppresses Airway Remodeling and Reduces Airway Hyperresponsiveness in an Animal Model. PLoS ONE, 11(7), e0158746.

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Cytokine Quantification in Cell Culture and Plasma

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For cytokine quantification in culture supernatants, the following ELISA kits have been used: Human/mouse TGF beta 1 2nd Generation ELISA Ready-SET-Go (eBioscience), Mouse IL-6 DuoSet ELISA (R&D Systems), Mouse IL-10 DuoSet ELISA (R&D Systems) and Mouse TNF alpha ELISA Ready-SET-Go (eBioscience) following the manufacturers' instructions. Cardiac troponin (cTnT) was quantified using Mouse cTnT/TNNT2 ELISA Kit (Immunoway).
A mouse multiplex Immunoassay (ProcartaPlex 4 Plex, Thermo Fisher) and Luminex 200 System (Thermo Fisher) were used to quantify cytokines in plasma samples.

Eghbalzadeh K., Georgi L., Louis T., Zhao H., Keser U., Weber C., Mollenhauer M., Conforti A., Wahlers T, & Paunel-Görgülü A. (2019). Compromised Anti-inflammatory Action of Neutrophil Extracellular Traps in PAD4-Deficient Mice Contributes to Aggravated Acute Inflammation After Myocardial Infarction. Frontiers in Immunology, 10, 2313.

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Multiplex Immunoassay for Cytokine Profiling

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A rat Multiplex Immunoassay (ProcartaPlex 4 Plex, Thermo Fisher) was used to quantify cytokines in plasma samples and a Simplex Kit (Rat ICAM-1 Procarta Plex Simplex, Thermo Fisher) for ICAM-1 quantification. All samples were analyzed using Luminex 200 System (Thermo Fisher).
Plasma concentrations of all other organ-specific injury markers were determined at the Central Institute of Clinical Chemistry and Laboratory Diagnostics of Düsseldorf University Hospital using automated analysers from Roche Diagnostics.

Weber C., Jenke A., Chobanova V., Yazdanyar M., Chekhoeva A., Eghbalzadeh K., Lichtenberg A., Wahlers T., Akhyari P, & Paunel-Görgülü A. (2019). Targeting of cell-free DNA by DNase I diminishes endothelial dysfunction and inflammation in a rat model of cardiopulmonary bypass. Scientific Reports, 9, 19249.

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Plasma Cytokine Profiling via Luminex

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EDTA plasma cytokine levels were determined by Luminex assay on Luminex 200 system using a 29-plex monkey panel according to the manufacturer’s instructions (LPC0005M; Thermo Fisher).

Bekerman E., Hesselgesser J., Carr B., Nagel M., Hung M., Wang A., Stapleton L., von Gegerfelt A., Elyard H.A., Lifson J.D, & Geleziunas R. (2019). PD-1 Blockade and TLR7 Activation Lack Therapeutic Benefit in Chronic Simian Immunodeficiency Virus-Infected Macaques on Antiretroviral Therapy. Antimicrobial Agents and Chemotherapy, 63(11), e01163-19.

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Multiplex Cytokine Analysis of BM Plasma

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BM plasma samples from the Mayo Clinic Rochester biobank were analyzed with Cytokine Human Magnetic 30-Plex panel for Luminex platform (LHC6003M, Life Technologies, Waltham, MA) and acquired on a Luminex 200 system equipped with with xPONENT 3.1 software (Thermo Fisher Scientific, Waltham, MA).

Calcinotto A., Brevi A., Chesi M., Ferrarese R., Garcia Perez L., Grioni M., Kumar S., Garbitt V.M., Sharik M.E., Henderson K.J., Tonon G., Tomura M., Miwa Y., Esplugues E., Flavell R.A., Huber S., Canducci F., Rajkumar V.S., Bergsagel P.L, & Bellone M. (2018). Microbiota-driven interleukin-17-producing cells and eosinophils synergize to accelerate multiple myeloma progression. Nature Communications, 9, 4832.

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Multiplex Cytokine Profiling of Mouse Plasma

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Mouse plasma samples, derived from cardiac puncture blood collected at euthanasia, were assayed for mouse GM-CSF, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-12, IL-13, IL-18, and TNF-α using a ProcartaPlex Mouse Th1/Th2 extended 11-plex kit (EXP110-20820-901; Thermo Fisher Scientific) on a Luminex 200 system (Thermo Fisher Scientific) according to the manufacturer’s protocol.

Viker K.B., Steele M.B., Iankov I.D., Concilio S.C., Ammayappan A., Bolon B., Jenks N.J., Goetz M.P., Panagioti E., Federspiel M.J., Liu M.C., Peng K.W, & Galanis E. (2022). Preclinical safety assessment of MV-s-NAP, a novel oncolytic measles virus strain armed with an H.pylori immunostimulatory bacterial transgene. Molecular Therapy. Methods & Clinical Development, 26, 532-546.

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HPV Genotyping Cervical Swab Protocol

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Sexual activities and vaginal medications were restricted prior to HPV analyses. Cervical discharges were swabbed for HPV detection, and genotyping was performed with a BioRad 100 Amplification and Luminex^® 200™ System (Thermo, Waltham, MA, USA) that detected 27 genotypes: high-risk genotypes 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, and 82 as well as low-risk genotypes 6, 11, 40, 42, 43, 44, 55, 61, 81, and 83.

Wei S., Wang K., Cheng F., Chang Z., Ren X., Liu Z., Liu M., Yang T., Ma X., Xie X, & Wang X. (2022). Prevalence of Human Papillomavirus Infection in the Female Partner of Infertile Couples Undergoing IVF/ICSI-ET and Subsequent Reproductive Outcomes. Journal of Clinical Medicine, 11(23), 7185.

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Cytokine Profiling in Murine Retinas

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Protein was extracted from the retinas from mice in each group at P17 with tissue lysate (Absin, Shanghai, China) containing protease inhibitors. The production of 8 cytokines (TNF-α, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-10, CXCL1) in retinal supernatants from each group was measured using the Mouse Cytokine Assay Kit (BIO-RAD, Richmond, VA, USA) according to the manufacturer’s instructions. The signals were detected and data analyzed by the Luminex 200 System (Thermo Scientific, San Diego, CA, USA).

Hong Y., Wang Y., Cui Y., Pan J., Mao S., Zhu Y., Wen T., Qi T., Wang A, & Luo Y. (2023). MicroRNA-124-3p Attenuated Retinal Neovascularization in Oxygen-Induced Retinopathy Mice by Inhibiting the Dysfunction of Retinal Neuroglial Cells through STAT3 Pathway. International Journal of Molecular Sciences, 24(14), 11767.

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Cytokine Profiling of Bone Marrow and Peripheral Blood in Multiple Myeloma

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Two femurs from each mouse were placed into a 0.5ml microtube with a hole at the bottom after their knee- caps were removed. The tube was placed inside a 1.5ml microtube, which was spun to obtained a BM cell pellet, which was suspended into 50ul PBS, and spun again. The obtained BM supernatant was stored frozen at −80°C till ready for analysis in duplicate with the Cytokine Mouse 20-Plex Panel for Luminex® Platform (LMC0006, Thermo Fisher Scientific) according to manufacturer’s instructions. Data were acquired on a Luminex200 system equipped with xPONENT 3.1 software (Thermo Fisher Scientific). Human BM plasma obtained from MM patients enrolled in the clinical trial at baseline and three or four weeks after LCL161 single agent treatment was assayed in duplicate using the Cytokine Human 30-Plex Panel for Luminex® Platform (LHC0003, Thermo Fisher Scientific). Similar analysis was performed on peripheral blood plasma from MM patients collected at baseline and four hours after LCL161 administration.

Chesi M., Mirza N.N., Garbitt V.M., Sharik M.E., Dueck A.C., Asmann Y.W., Akhmetzyanova I., Kosiorek H.E., Calcinotto A., Riggs D.L., Keane N., Ahmann G.J., Morrison K.M., Fonseca R., Lacy M.Q., Dingli D., Kumar S.K., Ailawadhi S., Dispenzieri A., Buadi F., Gertz M.A., Reeder C.B., Lin Y., Chanan-Khan A.A., Stewart A.K., Fooksman D, & Bergsagel P.L. (2016). IAP antagonists induce anti-tumor immunity in multiple myeloma. Nature medicine, 22(12), 1411-1420.

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