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Luminex xmap

Manufactured by Merck Group
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Sourced in United States, Germany

The Luminex xMAP is a multiplex technology platform that enables the simultaneous detection and quantification of multiple analytes in a single sample. It utilizes color-coded microspheres, flow cytometry, and proprietary signal detection to provide high-throughput, efficient, and flexible analysis capabilities.

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Close spelling variants of this product

LUMINEX xMAP MAGPIX Luminex xMAP-based immunoassays Luminex xMAP multiplexed bead system Luminex xMAP multianalyte profiling platform Milliplex™ MAP for Luminex® xMAP™ technology Milliplex xMAP Luminex Assay MAGPIX-Luminex XMAP technology Multiplex Biomarker Immunoassays for Luminex xMAP technology Luminex xMap platform multiplex assay Luminex xMAP instrument

16 protocols using luminex xmap

1

Quantification of Pioglitazone and Metabolites

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Plasma glucose was measured in the Clinical Research Center by the glucose oxidase method (Analox Glucose Analyzer, Analox Instruments, Lunenburg, MA). For other analyses, blood samples were placed on ice at the bedside, processed within 15–20 minutes, and frozen at −80°C until the samples were analyzed. Blood samples for the analysis of drug concentration were collected prior to the morning dose, and it was assured that patients were on chronic pioglitazone and that the drug concentration was at steady-state. A novel liquid chromatography tandem mass spectrometry assay described previously was used to measure plasma concentrations of pioglitazone, hydroxypioglitazone, and ketopioglitazone.17 The homeostasis model assessment of insulin resistance (HOMA-IR) was calculated from a single measurement of fasting insulin and glucose. Plasma insulin was determined by radioimmunoassay (Siemens, Los Angeles, CA). Adiponectin concentrations were measured by magnetic bead Milliplex technology (Luminex xMAP, Millipore, St. Charles, MO), and CK-18 levels were quantified by ELISA (M30-Apoptosense, DiaPharma, Columbus, OH).
Kawaguchi-Suzuki M., Bril F., Kalavalapalli S., Cusi K, & Frye R.F. (2017). Concentration-Dependent Response to Pioglitazone in Nonalcoholic Steatohepatitis. Alimentary pharmacology & therapeutics, 46(1), 56-61.
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2

Serum Biomarker Analysis Protocol

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The serum levels of Hcy and the transaminases alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured with an automated analyzer (Technicon RA-1000, Bayer Diagnostics, Leverkusen, Germany). Metallopeptidase inhibitor 1 (TIMP-1) was analyzed by the Luminex xMAP (Millipore, Darmstadt, Germany), a system that combines three basic forms of xMAP technology.
Gallego-Lopez M.D., Ojeda M.L., Romero-Herrera I., Nogales F, & Carreras O. (2022). Folic Acid Homeostasis and Its Pathways Related to Hepatic Oxidation in Adolescent Rats Exposed to Binge Drinking. Antioxidants, 11(2), 362.
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3

Comprehensive Mineral Metabolism Assessment

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All laboratory analyses were performed by standard techniques, specifically: ionic calcium [reference range (RR) = 4.6 to 5.3 mg/dl; selective ion method]; serum phosphate (RR = 2.7 to 4.5 mg/dl; enzymatic colorimetric method); iron (RR = 59 to 158 μg/dL; enzymatic colorimetric method); ferritin (RR = 30 to 400 ng/mL; eletroquimioluminescence method); transferrin saturation index (RR = 20 to 40%); alkaline phosphatase (RR = 32 to 122 U/L); parathyroid hormone (RR = 11 to 62 pg/ml, immunoradiometric assay); 25(OH)-vitamin D (RR = 30 to 100 ng/mL; quimioluminescence method); serum Fetuin-A (RR = 0.244 to 1,000 ng/mL; Luminex xMAP, Milliplex Analytes, Millipore Corp, St. Charles, MI). All samples were collected in fasting state.
Santos R.S., Coelho F.M., da Silva B.C., Graciolli F.G., Dominguez W.V., de Menezes Montenegro F.L., Jorgetti V., Moysés R.M, & Elias R.M. (2016). Parathyroidectomy Improves Restless Leg Syndrome in Patients on Hemodialysis. PLoS ONE, 11(5), e0155835.
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4

Quantification of Gut Hormones

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For GLP-1 measurement plasma was supplemented with aprotinin (1,000 KIU/ml) and dipeptidyl peptidase-4 inhibitor (DPP4–010; 20 μl/ml plasma; EMD Millipore, St. Charles, MO), for ghrelin it was acidified with 1 N hydrochloric acid (50 μl/ml) and treated with phenylmethanesulfonyl fluoride (0.1 mg/ml). Plasma insulin, c-peptide, glucagon, GIP, active GLP-1 (7–37 and 7–36 amide), peptide YY, pancreatic polypeptide and leptin were measured by multiplex immunoassay (Luminex xMAP, Millipore). Total ghrelin was determined by radioimmunoassay (Millipore). FGF-19 and FGF-21 were measured by enzyme-linked immunosorbent assay (R&D Systems, Minneapolis, MN).
Shibao C.A., Celedonio J.E., Tamboli R., Sidani R., Love-Gregory L., Pietka T., Xiong Y., Wei Y., Abumrad N.N., Abumrad N.A, & Flynn C.R. (2018). CD36 Modulates Fasting and Preabsorptive Hormone and Bile Acid Levels. The Journal of Clinical Endocrinology and Metabolism, 103(5), 1856-1866.
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5

Plasma Cytokine Profiling Protocol

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The blood samples of all the participants enrolled in this study were stored at 4 °C and centrifuged (2500× g, 10 min, 4 °C) within 2 h of collection. The harvested plasma samples were stored at −80 °C until analysis. A broad panel of cytokines was evaluated, including IFN-γ, IL-1β, IL-2, IL-6, IL-8, IL-10, IL-12p40, IL-12p70, TNF-α, monocyte chemoattractant protein (MCP) 1, and CRP. Fifty microliters of undiluted, freshly thawed plasma and twenty-five microliters of freshly thawed plasma diluted at 1:40,000 were used for the cytokine and CRP analyses, respectively. These samples were analyzed using a 96-well plate assay, as per the manufacturer’s instructions, using the Luminex® xMAP® magnetic bead platform (Milliplex Map Human Cytokine Panel; Millipore, Billerica, MA, USA). Standards provided by the manufacturer were assayed in duplicates to generate calibration lines in the range from 3.2 to 10,000 pg/mL for each cytokine and from 0.01 to 50 ng/mL. The coefficients of variation and the relative standard errors of the standard curve and quality control samples were <15%.
Benzi J.R., Melli P.P., Duarte G., Unadkat J.D, & Lanchote V.L. (2023). The Impact of Inflammation on the In Vivo Activity of the Renal Transporters OAT1/3 in Pregnant Women Diagnosed with Acute Pyelonephritis. Pharmaceutics, 15(10), 2427.
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6

Endocrine and Metabolic Biomarkers

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Corticosterone, progesterone and testosterone levels were determined with ALPCO assay kits (Salem, NH, USA). Adrenocorticotropic hormone (ACTH) levels were measured using an ELISA kit from G Biosciences (St. Louis, MO, USA). LiquiColor kits for assay of plasma triglyceride and total cholesterol levels analysis were obtained from Stanbio Laboratory (Boerne, TX, USA). Free-fatty acids (FFA) levels were determined with kits from Wako Diagnostic (Richmond, VA, USA). Serum obtained from mice was analyzed for adipokines (including IL-6, insulin, leptin, MCP-1, PAI-1, resistin, and TNF-α) using a multiplex mouse adipokines assay (EZMADP-60K from Millipore, Bedford, MA, USA), and detected by Luminex xMAP (Millipore).
Hao X., Zhu B., Yang P., Dong D., Sahbaie P., Oliver P.L., Shen W.J., Azhar S, & Kraemer F.B. (2021). SNAP25 mutation disrupts metabolic homeostasis, steroid hormone production and central neurobehavior. Biochimica et biophysica acta. Molecular basis of disease, 1868(2), 166304.
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7

Comprehensive Metabolic Biomarker Profiling

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Glucose, total NEFA TG, plasma cortisol, TSH, free T3 and free T4 were measured using specific radioimmunoassays and colorimetric assays (Ouellet et al., 2012) . Plasma glucagon, insulin and leptin were measured using Luminex xMAP-based immunoassays (Millipore, Etobicoke, ON, Canada). Adiponectin, total and acylated ghrelin were measured by ELISA (Alpco, Salem, NH, USA). Individual plasma NEFA (palmitate, linoleate, oleate), [U-13 C]palmitate enrichment and [1,1,2,3,3-2 H]glycerol enrichment were measured by gas chromatography-mass spectrometry (Ouellet et al., 2012) . [3-3 H]-glucose specific activity was determined by liquid scintillation spectrometry (Ouellet et al., 2012) .
Blondin D.P., Nielsen S., Kuipers E.N., Severinsen M.C., Jensen V.H., Miard S., Jespersen N.Z., Kooijman S., Boon M.R., Fortin M., Phoenix S., Frisch F., Guérin B., Turcotte É.E., Haman F., Richard D., Picard F., Rensen P.C.N., Scheele C, & Carpentier A.C. (2020). Human Brown Adipocyte Thermogenesis Is Driven by β2-AR Stimulation. Cell metabolism, 32(2).
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8

Comprehensive Protein Profiling of Biological Samples

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The plasma protein concentrations of ADAMTS13, eotaxin, Fas, GDF15, GROα, ICAM1, IL1α, IL6, IL7, IL8, IL10, IL15, MCP1, MDC, MIP1α, MIP1β, MMP1, MMP7, MMP9, MPO, OPN, PAI1, PARC, RAGE, RANTES, SOST, TARC, TNFR1, TNFR2, TNFα, and VEGFA were quantified using commercially available multiplex magnetic bead-based immunoassays (R&D Systems) on the Luminex xMAP multianalyte profiling platform and analyzed on MAGPIX System (Merck Millipore). Activin A concentration was determined by a Quantikine ELISA Kit (R&D Systems). Protein names, abbreviations, and aliases are provided in Additional file 1: Table S1. All assays were performed according to the manufacturer’s protocols. In instances where a biomarker was below the limit of detection in a participant sample, a value of half of the lowest measured value for that analyte was assigned.
Aversa Z., Atkinson E.J., Carmona E.M., White T.A., Heeren A.A., Jachim S.K., Zhang X., Cummings S.R., Chiarella S.E., Limper A.H, & LeBrasseur N.K. (2023). Biomarkers of cellular senescence in idiopathic pulmonary fibrosis. Respiratory Research, 24, 101.
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9

Multiplex Biomarker Profiling in Plasma

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Soluble biomarkers were measured using multiplex magnetic bead immunoassay based on Luminex® xMAP multianalyte profiling platform and analyzed on MAGPIX® System (Merck Millipore) at the Institute of Medical Biochemistry and Laboratory Diagnostics of The General University Hospital and of the First Faculty of Medicine of Charles University in Prague. The following premixed magnetic bead kits were used for plasma sample analysis: LXSAHM-07: endoglin (BR22), IL-1β (BR28), IL-22 (BR35), IL-6 (BR13), procalcitonin (BR39), VCAM-1 (BR57), and VEGF-A (BR26) and LXSAHM-08: angiopoietin-2 (BR26), endothelin-1 (BR76), ICAM-1 (BR61), IL-1α (BR38), IL-17A (BR42), IL-18 (BR78), IL12 p70 (BR563), TNF-α (BR12) (both from Biotechne R&D Systems), and Milliplex MAP human soluble receptor magnetic bead panel (HSCRMAG-32K-02: VEGFR1 and VEGFR2) and cardiovascular disease (CVD) magnetic bead panel (HCVD1MAG-67K-01: endocan1/ESM1) (Merck).
Vítková V., Pánek M., Janec P., Šibíková M., Vobruba V., Haluzík M., Živný J, & Janota J. (2018). Endothelial Microvesicles and Soluble Markers of Endothelial Injury in Critically Ill Newborns. Mediators of Inflammation, 2018, 1975056.
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10

Multiplex Cytokine Analysis in Serum

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Unbiased analysis of serum cytokines was performed using Luminex xMAP multiplex technology and the MILLIPLEX MAP Human Cytokine/Chemokine Magnetic Bead Panel Kit, according to the manufacturer’s standard protocol (Merck, Rahway, NJ, USA). Data processing was carried out using Belysa software v1.1.0 (Merck, Rahway, NJ, USA) at the Resource Center “Cell Technology and Immunology”, Sirius University of Science and Technology.
Vavilova J.D., Ustiuzhanina M.O., Boyko A.A., Streltsova M.A., Kust S.A., Kanevskiy L.M., Iskhakov R.N., Sapozhnikov A.M., Gubernatorova E.O., Drutskaya M.S., Bychinin M.V., Novikova O.N., Sotnikova A.G., Yusubalieva G.M., Baklaushev V.P, & Kovalenko E.I. (2023). Alterations in the CD56− and CD56+ T Cell Subsets during COVID-19. International Journal of Molecular Sciences, 24(10), 9047.
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