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Luminex 200 analyzer

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The Luminex 200 analyzer is a multiplex system that utilizes flow cytometry technology to simultaneously detect and quantify multiple analytes in a single sample. The instrument is capable of analyzing up to 100 different targets in a single reaction well.

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Lab products found in correlation

Milliplex map kit, by Merck Group (6 mentions) Multiplex fluorescent bead based immunoassays, by DiaSorin (4 mentions) Human cytokine chemokine magnetic bead panel, by Merck Group (4 mentions) Xponent software, by DiaSorin (3 mentions) Elisa kit, by R&D Systems (3 mentions) Analyst software, by Merck Group (2 mentions) Milliplex analyst software, by Merck Group (2 mentions) Vacutainer plus, by BD (2 mentions) Human cytokine antibody immobilized magnetic beads, by Merck Group (2 mentions) Microplate spectrophotometer, by Bio-Rad (2 mentions) Bio plex manager software, by Bio-Rad (2 mentions) Magnetic bead suspension assay, by Merck Group (2 mentions) Luminex kit, by R&D Systems (2 mentions) Luminex assay, by Thermo Fisher Scientific (1 mentions) Dounce homogenizer, by Thomas Scientific (1 mentions) Fuji dri chem 7000v, by Fujifilm (1 mentions) Milliplex map, by Merck Group (1 mentions) Milliplex map system, by Merck Group (1 mentions) Luminex analyzer, by DiaSorin (1 mentions) Bio plex pro mouse cytokine panel 33 plex, by Bio-Rad (1 mentions)

85 protocols using luminex 200 analyzer

1

Inflammatory Cytokine Profiling of Athlete-Derived Sera

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After growing 3T3-L1 and C2C12 cells for 72 h under the four culture conditions (supplementation with 10% serum from HP, HE, and LP/LE athletes or non-athlete control), media were collected to measure the concentration of secreted (mouse-specific) cytokines from the treated cells in order to evaluate the inflammatory status triggered by these interventions. Accumulated levels of six secreted mice cytokines (IFN gamma, IL-1 beta, IL-6, IL-10, IL-17A (CTLA-8), and TNF-alpha) in the media supernatants were measured using the inflammatory cytokine ProcartaPlex™ mouse and rat mix and match 6-plex panels (PPX-06-MXXGTDN, Thermo Fisher Scientific) using the Luminex™ 200 analyzer according to the manufacturer’s instructions (Luminex, Madison, WI, United States). Data were analyzed using xPONENT 4.2 software (Luminex™ 200 analyzer, Luminex, Madison, WI, United States).
Alheidous S., Al-Muraikhy S., Rizk N., Sellami M., Donati F., Botre F., Al-Mansoori L, & Elrayess M.A. (2022). Effect of sera from elite athletes on cytokine secretion and insulin signaling in preadipocytes and skeletal muscle cells. Frontiers in Molecular Biosciences, 9, 943034.
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2

Cytokine Quantification from Cell Supernatants

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Supernatants were obtained from stimulated single-cell suspensions (SPN, MLN, CLN, LPL, and EYE), and stored at −80°C until the Luminex assay (Life Technologies) was performed, after which cytokine production was quantified through a Luminex 200 analyzer (Luminex Corp., Austin, TX, USA).
Llorenç V., Nakamura Y., Metea C., Karstens L., Molins B, & Lin P. (2022). Antimetabolite Drugs Exhibit Distinctive Immunomodulatory Mechanisms and Effects on the Intestinal Microbiota in Experimental Autoimmune Uveitis. Investigative Ophthalmology & Visual Science, 63(3), 30.
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3

Cytokine and Chemokine Profiling after SCI

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The blood from individual animals was collected 12 h after SCI into EDTA tubes to obtain plasma. Frozen samples of isolated spinal cord were homogenized (Dounce homogenizer, Thomas Scientific, Swedesboro, NJ, USA) and transferred to 10 volumes (w/w) of ice-cold PBS supplemented with 0.1% Tween 20 and a protease inhibitor cocktail. The prepared spinal cord homogenate was vortexed for 15 s and subjected to rotator for 5 min at room temperature. Further cell debris was removed by centrifugations at 4 °C (30,000× g for 10 min). The supernatant was further used for measurement. Milliplex Rat Cytokine/Chemokine Magnetic Bead Panel (Merck, Rahway, NJ, USA) was used to measure the levels of chemokines and cytokines in plasma and spinal cord extracts. Samples were diluted at 1:4 ratio and further incubated with magnetic beads, washed up, and then incubated with detecting antibodies and SA-PE. The data were obtained using a Luminex 200 analyzer using xPONENT software (Luminex, Austin, TX, USA).
Telegin G.B., Chernov A.S., Minakov A.N., Rodionov M.V., Kazakov V.A., Palikov V.A., Balmasova I.P., Asyutin D.S., Poluektov Y.M., Konovalov N.A., Kudriaeva A.A., Spallone A., Gabibov A.G, & Belogurov AA J.r. (2022). Plasma Cytokines Level and Spinal Cord MRI Predict Clinical Outcome in a Rat Glial Scar Cryoinjury Model. Biomedicines, 10(10), 2345.
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4

Multiplex Cytokine Profiling via Luminex 200 Analyzer

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Luminex 200 analyzer (Luminex Corp, Austin, TX) was used to detect the cytokines, chemokines, and growth factors in the serum using custom-made 48-plex multiplex kits (Bio-Rad Laboratories, Hercules, CA). A list of these 48 cytokines has been included in the supplementary file (https://doi.org/10.1667/RADE-22-001011.1.S1). Standard curves for each cytokine were prepared by serial dilution and run-in duplicates. Cytokine concentration (pg/ml) was determined by fluorescence intensity and its quantification was performed using Bio-Plex Manager software, version 6.1 (Bio-Rad Inc.) (35 (link)).
Zhao Y., Guo Y.J., Tomac A.C., Taylor N.R., Grinberg A., Lee E.J., Huang S, & Westphal H. (1999). Isolated cleft palate in mice with a targeted mutation of the LIM homeobox gene Lhx8. Proceedings of the National Academy of Sciences of the United States of America, 96(26), 15002-15006.
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5

Quantifying Insulin Signaling Biomarkers

Cited 2 times
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The change in phosphorylation of insulin signaling biomarkers [p70S6K (Thr412), IRS1 (Ser636), GSK3-α (Ser21), GSK3-β (Ser9), Akt (Ser473), PTEN (Ser380), IR (Tyr1162/Tyr1163), IGF1R (Tyr1135/Tyr1136), RPS6 (Ser235/Ser236), TSC2 (Ser939), and mTOR (Ser2448)] was quantified in lysates containing equal concentration of proteins (25 μg) prepared from 3T3-L1 and C2C12 cells using an Akt/mTOR phosphoprotein 11-plex magnetic bead kit 96-well plate (48-611MAG, Millipore MILLIPLEX, United States), following the manufacturer’s instructions. Mean fluorescent intensity (MFI) was assessed using Luminex 200 using xPONENT 4.2 software (Luminex™ 200 analyzer, Luminex, Madison, WI, United States). Table 1 lists these proteins and summarizes their roles in insulin signaling (Gual et al., 2005 (link); Jolivalt et al., 2008 (link); Frosig and Richter, 2009 (link); Zhang et al., 2009 (link)).
Alheidous S., Al-Muraikhy S., Rizk N., Sellami M., Donati F., Botre F., Al-Mansoori L, & Elrayess M.A. (2022). Effect of sera from elite athletes on cytokine secretion and insulin signaling in preadipocytes and skeletal muscle cells. Frontiers in Molecular Biosciences, 9, 943034.
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6

Microsphere-based Multiplex Assay

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The PCR products were conjugated with MagPlex-TAG microspheres, which were pre-coupled with “anti-TAG” sequence. The working mixture containing 2500 of each target microspheres was diluted with 1× Tm Hybridization Buffer (0.2 M NaCl, 0.1 M Tris, 0.08% Triton X-100, Ph 8.0, filter sterilized). For each reaction, 5 μl of amplified product or distilled water, 75 μl of SAPE solution and 20 μl of the working MagPlex-TAG microsphere mixture were well mixed together before incubated in a thermocycler for 30 min at 45 °C. The Luminex 200 analyzer was applied to analyze the products after the hybridization reaction.
Wu M., Zhu Y., Cong F., Rao D., Yuan W., Wang J., Huang B., Lian Y., Zhang Y., Huang R, & Guo P. (2018). Rapid detection of three rabbit pathogens by use of the Luminex x-TAG assay. BMC Veterinary Research, 14, 127.
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7

Multiplex Luminex Immunoassays for Neurotrophic Factors

Cited 1 time
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Concentrations of selected neurotrophic factors (NT-3, NGF, BDNF) and CRP were assessed in CSF and plasma using multiplex fluorescent bead-based immunoassays (Luminex Corporation, TX, USA). Assays were performed according to manufacturer’s protocol, as described previously [79 (link)]. Final concentration of the analyzed protein was assessed based on the averaged readings (average %CV = 6.8) obtained from both duplicates of a given sample in relation to four (one for each of the analytes) 6-point standard curves showing median fluorescence intensity. Before each run, the Luminex 200 analyzer was calibrated, and the performance verification was performed according to manufacturer’s instructions.
Baumert B., Sobuś A., Gołąb-Janowska M., Ulańczyk Z., Paczkowska E., Łuczkowska K., Zawiślak A., Milczarek S., Osękowska B., Meller A., Machowska-Sempruch K., Wełnicka A., Safranow K., Nowacki P, & Machaliński B. (2020). Local and Systemic Humoral Response to Autologous Lineage-Negative Cells Intrathecal Administration in ALS Patients. International Journal of Molecular Sciences, 21(3), 1070.
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8

Comprehensive Serum Biomarker Analysis

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Serum glucose (GLU), triglyceride (TG), glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), alkaline phosphatase (ALP), high-density lipoprotein (HDL) and total cholesterol (TCHO) were measured using a serum chemistry analyzer (Fuji Dri-chem 7000V, FujiFilm corporation, Tokyo, Japan). Serum leptin and resistin were analyzed with a Milliplex MAP kit (Merck Millipore, Burlington, MA, USA) using a Luminex 200 analyzer (Luminex, Austin, TX, USA).
Kanno R., Koshizuka T., Miyazaki N., Kobayashi T., Ishioka K., Ozaki C., Chiba H, & Suzutani T. (2021). Protection of Fatty Liver by the Intake of Fermented Soybean Paste, Miso, and Its Pre-Fermented Mixture. Foods, 10(2), 291.
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9

Multiplex Cytokine Quantification

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A Milliplex Map kit (EMD Millipore) was used to measure seven cytokines (on previously frozen samples) in one assay (IL-1α, IL-1β, IL-4, IL-6, IFNγ, IL-8, and TNF-α). Plates were read in a Luminex 200 Analyzer (Luminex) controlled by xPONENT software. Values for each analyte were determined using Analyst software (EMD Millipore) from a standard curve of log dose versus median fluorescent intensity using a 5-parameter logistic fit.
Balbuena-Merle R., Curtis S.A., Devine L., Gibb D.R., Karafin M.S., Luckey C.J., Tormey C.A., Siddon A.J., Roberts J.D, & Hendrickson J.E. (2019). Red blood cell alloimmunization is associated with lower expression of FcγR1 on monocyte subsets in patients with sickle cell disease. Transfusion, 59(10), 3219-3227.
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10

Multiplex Serology for HPV and HPyV Antibodies

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Baseline peripheral blood samples were collected, and serum aliquots were stored at −70°C until further processing at the German Cancer Research Center. Serum samples were analyzed for antibodies to the major capsid protein L1 corresponding to 17 beta‐HPV types [species 1 (types 5, 8, 12, 21, 24, 36, 93), species 2 (types 9, 15, 17, 22, 23, 38, 80), species 3 (type 75), species 4 (type 92) and species 5 (type 96)], seven gamma‐HPV types (4, 48, 50, 60, 88, 101 and 103) and four HPyV types (6, 7, trichodysplasia spinulosa‐associated polyomavirus and Merkel cell polyomavirus [MCPyV]). Multiplex serological assays were used to allow for measurement of multiple antibodies in one cycle. The antibody detection method was based on Glutathione S‐Transferase capture enzyme‐linked immunosorbent assay38, 39 in combination with Luminex fluorescent bead technology,24, 40, 41 as described in detail previously. A Luminex 200 analyzer (Luminex Corp., Austin, Texas) was used to identify the internal color of the individual beads and quantify their reporter fluorescence, expressed as median fluorescence intensity.24, 40, 41
Zhao Y., Amorrortu R.P., Fenske N.A., Cherpelis B., Messina J.L., Sondak V.K., Giuliano A.R., Schell M.J., Waterboer T., Pawlita M., McKay‐Chopin S., Gheit T., Tommasino M, & Rollison D.E. (2020). Cutaneous viral infections associated with ultraviolet radiation exposure. International Journal of Cancer, 148(2), 448-458.
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