Bio plex 200 luminex instrument

Manufactured by Bio-Rad

Sourced in United States, Sweden

The Bio-Plex 200 Luminex instrument is a multiplex assay system that uses the Luminex xMAP technology. It is capable of simultaneously detecting and quantifying multiple target analytes in a single small-volume sample.

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Bio-Plex 200 (546 citations) Bio-Plex (286 citations) Bio-Plex 200 instrument (124 citations) Luminex 200 (47 citations) Bio-Plex instrument (16 citations) Luminex 200 instrument (13 citations) Bio-Plex (Luminex 200) (10 citations) Luminex instrument (7 citations) Luminex Bio-Plex 200 IS 100 instrument (2 citations) Bio-Plex Luminex 200 XYP instrument (2 citations)

16 protocols using bio plex 200 luminex instrument

Multiplex Cytokine Analysis in FINGER Study

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In the FINGER study, venous blood samples were taken at baseline in fasting status and using EDTA tubes. Plasma aliquots were stored at – 80 °C until analysis. A panel of cytokines, chemokines, and growth factors (IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12(p40), IL-12(p70), IL-13, IL-15, IL-17, IFN-α2, IFN-γ, MIP-1α, TNF-β) were analyzed with a multiplex suspension array system using Bioplex Luminex 200 instrument (Bio-Rad Laboratories, Hercules, CA, USA) and the MILLIPLEX® MAP Human Cytokine/Chemokine panel (Merck Millipore, Darmstadt, Germany).
The assays were performed in one batch, and samples preparation and setting of the system running protocol were done following the manufacturer’s instructions. All samples and standards were run in duplicate and were measured as pg/ml. Quality controls were performed according to the manufacturer guidelines to ensure accuracy of measurements. After the plate reading, the results files were generated using Bio-Plex Manager software 4 (Bio-Rad Laboratories, Hercules, CA, USA).

Goikolea J., Gerenu G., Daniilidou M., Mangialasche F., Mecocci P., Ngandu T., Rinne J., Solomon A., Kivipelto M., Cedazo-Minguez A., Sandebring-Matton A, & Maioli S. (2022). Serum Thioredoxin-80 is associated with age, ApoE4, and neuropathological biomarkers in Alzheimer’s disease: a potential early sign of AD. Alzheimer's Research & Therapy, 14, 37.

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Cytokine Profiling in Virus Infection

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The concentrations of six core cytokines in virus-infected culture supernatants were determined using the Bio-Plex Pro Human Cytokine 27-plex panel (Bio-Rad Laboratories). Specifically, the following analytes were included for statistical analysis: IL-1β, IL-6, IL-8, IFN-γ, IP-10, and TNF-α. Samples were initially incubated on antibody-coupled beads for 60 min to allow binding, followed by incubation with detection antibodies for 30 min. Conjugates were treated with streptavidin for 10 min, washed on a Bio-Plex Pro II wash station, resuspended, vortexed, and quantified by fluorescence. All data were analyzed using a Bio-Rad Bio-Plex Luminex 200 instrument. Standard curves (Log (x) – Linear (y)) were generated with the Bio-Rad Bio-Plex Manager v6.0 software. Triplicate measurements of all samples were performed.

Huang C.G., Lee L.A., Wu Y.C., Hsiao M.J., Horng J.T., Kuo R.L., Huang C.H., Lin Y.C., Tsao K.C., Chen M.C., Chen T.C, & Shih S.R. (2018). A pilot study on primary cultures of human respiratory tract epithelial cells to predict patients’ responses to H7N9 infection. Oncotarget, 9(18), 14492-14508.

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Multiplex Cytokine Profiling in Systemic Inflammation

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Systemic inflammation biomarkers, including adaptive immunity cytokines, proinflammatory cytokines, and anti-inflammatory cytokines, were measured using a single multiplex kit (Bio-Plex Pro Human Cytokine 27-plex panel, Bio-Rad Laboratories, Hercules, CA, USA). Patients with acute systemic infections or inflammation were not tested until the conditions had subsided. 25 Whole blood was obtained in the morning, and serum was separated from each sample. The serum specimens were immediately stored at -80°C until assay. The commercial kit allowed for the measurement of concentrations of 27 cytokines and chemokines, including IL-1β, IL-1 receptor antagonist (IL-1ra), IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12, IL-13, IL-15, IL-17, eotaxin, basic fibroblast growth factor (basic-FGF), eotaxin, granulocyte-colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon-γ, interferon gamma-induced protein-10 (IP-10), monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein (MIP)-1α, MIP-1β, platelet-derived growth factor-BB (PDGF-BB), CCR5, TNF-α, and vascular endothelial growth factor. All of the procedures followed the recommendations of the manufacturer. Duplicate measurements of all samples were assessed using a Bio-Rad Bio-Plex Luminex 200 instrument and Bio-Rad Bio-Plex Manager software (v6.0).

Huang C.G., Hsu J.F., Chuang L.P., Li H.Y., Fang T.J., Huang Y.S., Yang A.C., Lee G.S., Kuo T.B.J., Yang C.C.H., Lee L.A, & Chuang H.H. (2023). Adenotonsillectomy-related changes in systemic inflammation among children with obstructive sleep apnea. Journal of the Chinese Medical Association : JCMA, 86(6).

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Luminex Bead-Based Multiplex Immunoassay

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Coupling Luminex MagPlex-COOH beads (Bio-Rad; Hercules, CA) to monoclonal antibodies was based on the procedures outlined in the X-MAP Cookbook [15 ]. All primary antibodies that contained amine containing additives or preservatives were cleaned using Micro Bio-Spin 6 Tris chromatography columns (Bio-Rad; Hercules, CA) according to the manufacturer’s instructions. Table 2 lists the antibodies that were conjugated to MagPlex beads to detect Aβ42, pTau, GFAP and IBA1.
Conjugation of antibodies to Luminex™ MagPlex-COOH beads was performed using the Bio-Rad bead making kit (Bio-Rad; Hercules, CA), and conjugated beads were quantified using a TC20 cell counter (Bio-Rad; Hercules, CA). For confirmation of antibody coupling to beads, serial dilutions of phycoerythrin (PE) labeled anti-species IgG detection antibodies (Jackson ImmunoResearch; West Grove, PA) were used to detect primary bead-coupled antibodies from mouse or rabbit hosts (Table 2). The beads were then read using the Bio-Plex® 200 Luminex instrument (Bio-Rad; Hercules, CA).

Keene C.D., Wilson A.M., Kilgore M.D., Bruner L.T., Postupna N.O, & Darvas M. (2018). Luminex-based quantification of Alzheimer’s Disease neuropathologic change in formalin-fixed post-mortem human brain tissue. Laboratory investigation; a journal of technical methods and pathology, 99(7), 1056-1067.

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Luminex Bead-Based Multiplex Immunoassay

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Multiplexed CSF Biomarker Quantification

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CSF samples were collected via lumbar puncture. Biomarker assays were performed using multiplexed (Luminex platform) immunometric assays (R & D Systems, Minneapolis, Minnesota, USA) according to manufacturer directions, using a Bio-Plex 200 Luminex instrument and Bio-Plex analysis software (Bio-Rad, Hercules, California, USA). CSF levels of CCL2, IL-2, IL-6,TNF-α, IFN-γ, soluble tumor necrosis factor receptor 2 (sTNFR2), sIL-6Rα, sIL-2, sCD14 and B-cell activating factor (BAFF) were quantified. Of note, due to budgetary constraints, we did not quantify inflammatory markers in plasma; however, studies have shown that CSF inflammatory markers are highly correlated with plasma [26 (link),27 (link)].

Thames A.D., Briones M.S., Magpantay L.I., Martinez-Maza O., Singer E.J., Hinkin C.H., Morgello S., Gelman B.B., Moore D.J., Heizerling K, & Levine A.J. (2015). The role of chemokine C-C motif ligand 2 genotype and cerebrospinal fluid chemokine C-C motif ligand 2 in neurocognition among HIV-infected patients. AIDS (London, England), 29(12), 1483-1491.

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Multiplex Cytokine Profiling in Serum

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The serum concentrations of cytokines, chemokines, and growth factors were analyzed from unthawed serum samples with Luminex technology using the 38-plexed Milliplex MAP Kit (cat.no. HCYTMAG-60K-PX38) according to the manufacturer’s recommendations (Merck-Millipore Corp., Billerica, MA, USA). Luminex analyses were performed with single reactions using undiluted serum samples. Quantification of the markers was performed with Bio-plex 200 Luminex instrument and Bio-Plex Manager software (Bio-Rad, Sweden). The concentration of each marker was determined from an 8-point standard curve using five-parameter logistic regression. Minimum detectable concentration (MinDC) was determined for each marker separately using the lowest concentration on the standard curve linear phase [MinDC=c(low)+2SD]. The samples below MinDC were given a value of 50% of the MinDC. We categorized the cytokines into six groups: Th1, Th2, Th17, and Treg cytokines; chemokines; growth factors; innate immune cytokines; other cytokines; and other inflammation-related markers (Figure 1).

Niinistö S., Miettinen M.E., Cuthbertson D., Honkanen J., Hakola L., Autio R., Erlund I., Arohonka P., Vuorela A., Härkönen T., Hyöty H., Krischer J.P., Vaarala O., Knip M, & Virtanen S.M. (2022). Associations Between Serum Fatty Acids and Immunological Markers in Children Developing Islet Autoimmunity—The TRIGR Nested Case–Control Study. Frontiers in Immunology, 13, 858875.

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Multiplex Cytokine and Chemokine Profiling

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Two electrochemiluminesence-based multiplex assay panels (Proinflammatory 9-plex and Chemokine 7-plex; Meso-Scale Diagnostics, LLC, Rockville, MD) were used to determine concentrations of IL-1β, IL-2, IL-6, IL-10, IL-12p70, GM-CSF, IFN-γ, TNF-α, CXCL8 (IL-8), CXCL10 (IP-10), CCL11 (eotaxin), CCL2 (MCP-1), CCL13 (MCP-4), CCL4 (MIP-1β), and CCL17 (TARC). All testing was done at a centralized laboratory. Analyte- and plate-specific lower limits of detection (LLOD) were calculated as concentrations 2.5 standard deviations above the background for each analyte on each plate. Concentrations of five soluble receptors (sCD14, sgp130, sIL-2Rα, sTNF-R2), a cytokine (BAFF), and a chemokine CXCL13 (BLC-BCA1), were measured in a single panel (Human Biomarker Custom Premix Kit A) using the fluorescent bead-based multiplexed Luminex xMAP system at a centralized laboratory (Fluorokine^® MAP, R&D Systems, Minneapolis, MN), and a Bio-Plex 200 Luminex instrument and Bio-Plex software (Bio-Rad, Hercules, CA). A single assay lot was used. Finally, CRP was measured at Quest Diagnostics using a high-sensitivity nephelometric assay (Dade Behring, Inc., Newark, DE). All specimens for any given individual were run on the same plate.

McKay H.S., Bream J.H., Margolick J.B., Martínez-Maza O., Phair J.P., Rinaldo C.R., Abraham A.G, & Jacobson L.P. (2016). Host factors associated with serologic inflammatory markers assessed using multiplex assays. Cytokine, 85, 71-79.

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Multiplex Cytokine Quantification

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The serum concentrations of IFNG, IL-17A and IL-22 were analyzed using the Milliplex MAP Kit (HTH17MAG-14K) according to the manufacturer's recommendations (Merck-Millipore Corp., Billerica, MA, USA). Quantification of the markers was performed with a Bio-plex 200 Luminex-instrument and Bio-Plex Manager software (Bio-Rad, Sweden). The samples below Minimum detectable concentration (MinDC) DC were given an arbitrary value of 50% of MinDC.

Honkanen J., Vuorela A., Muthas D., Orivuori L., Luopajärvi K., Tejesvi M.V., Lavrinienko A., Pirttilä A.M., Fogarty C.L., Härkönen T., Ilonen J., Ruohtula T., Knip M., Koskimäki J.J, & Vaarala O. (2020). Fungal Dysbiosis and Intestinal Inflammation in Children With Beta-Cell Autoimmunity. Frontiers in Immunology, 11, 468.

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Multiplex Cytokine Quantification from Serum Samples

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Serum concentrations of cytokines, chemokines, and growth factors (listed in Supplementary Table 6) were analyzed using the 38-plexed Milliplex MAP Kit (cat.no. HCYTMAG-60K-PX38) according to the manufacturer's recommendations (Merck-Millipore Corp., Billerica, MA, USA). Analyses were performed in single reactions. Quantification of the markers was performed with a Bio-Plex 200 Luminex instrument and Bio-Plex Manager software (Bio-Rad, Sweden). Concentration of each marker was determined from an 8-point standard curve using five parameter logistic regression. Minimum detectable concentration (MinDC) was determined for each marker separately using the lowest concentration on the standard curve's linear phase [MinDC = c(low) + 2SD where SD is standard deviation]. The samples below MinDC were given a value of 50% of MinDC. In total, 38 different cytokines were isolated and quantified from 30 representative samples (n = 20 diet group, n = 10 control group) gathered from the study population.

Sarin H.V., Gudelj I., Honkanen J., Ihalainen J.K., Vuorela A., Lee J.H., Jin Z., Terwilliger J.D., Isola V., Ahtiainen J.P., Häkkinen K., Jurić J., Lauc G., Kristiansson K., Hulmi J.J, & Perola M. (2019). Molecular Pathways Mediating Immunosuppression in Response to Prolonged Intensive Physical Training, Low-Energy Availability, and Intensive Weight Loss. Frontiers in Immunology, 10, 907.

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