Luminex assay

Manufactured by Merck Group

Sourced in United States, Germany, Sweden

The Luminex assay is a multiplex technology that enables the simultaneous detection and quantification of multiple analytes in a single sample. It utilizes color-coded magnetic beads coated with capture antibodies specific to different target proteins or molecules. The analytes in the sample bind to their respective capture antibodies, and the resulting complexes are then detected and quantified using a specialized flow-based detection system.

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Close spelling variants of this product

Milliplex Multiplex Assays Luminex xMAP Milliplex MAP Luminex microbead assays Multiplex Fluorescent Bead-Based Luminex Cytokine Assays Luminex bead array assay Luminex assay for 17 human cytokines Luminex bead assay Mouse Cytokine/Chemokine Luminex 25plex Assay Luminex Human isotyping assay Luminex multiplex assay Luminex bead-based assay

50 protocols using luminex assay

Radiation-Induced Hormonal and Inflammatory Changes

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Blood samples were collected from the tail in the morning around 10 am and placed at room temperature for about 60 minutes to coagulate and then centrifuged at 1000 × g for 10 minutes at 5 and 20 weeks after irradiation. The serum was then removed and stored at −80°C. A Luminex assay was used according to the manufacturer's instructions (Millipore, Stockholm, Sweden) to detect pituitary‐related hormones (MILLIPLEX MAP rat pituitary magnetic bead panel assay kit). Reproduction‐related hormone analyses were performed using ultrasensitive rat estradiol, testosterone and progesterone ELISA kits (Crystal Chem, IL, USA). To compare pro‐inflammatory mediator expression profiles between irradiated and non‐irradiated control rats, a Luminex assay was used according to the manufacturer's instructions (Millipore, Stockholm, Sweden). The results were analysed using a Bio‐Plex workstation (Bio‐Rad, Hercules, CA, USA) and normalized to the amount of total protein extracted from hypothalamus homogenates using a colorimetric Bio‐Rad DC Protein Assay kit (Bio‐Rad).

Xu Y., Sun Y., Zhou K., Xie C., Li T., Wang Y., Zhang Y., Rodriguez J., Zhang X., Shao R., Wang X, & Zhu C. (2020). Cranial irradiation alters neuroinflammation and neural proliferation in the pituitary gland and induces late‐onset hormone deficiency. Journal of Cellular and Molecular Medicine, 24(24), 14571-14582.

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Biomarker Analysis of CPI Treatment

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The KUCC BRCF protocol permits collection of up to 6 tubes of blood from each participant at each visit. Lab sampling occurred during scheduled venipuncture for standard of care lab sampling in conjunction with participants’ CPI treatment at baseline and six months. These samples were de-identified (marked only with participants’ study identification numbers) and stored until samples from all timepoints from all participants were collected. At that time, the KUCC Biomarker Discovery Lab (BDL) staff obtained 1 mL of the deidentified plasma baseline and 6-month samples to quantify circulating levels of the study biomarkers using Luminex assays (Millipore-Sigma, Temecula, CA, USA) analyzed on a BioPlex 200 instrument. The mean-fluorescent intensity data generated by the Luminex assays for each marker was compared to standard curves and the resulting concentrations were supplied to the biostatistician for the analyses described below.

Myers J.S., Parks A.C., Mahnken J.D., Young K.J., Pathak H.B., Puri R.V., Unrein A., Switzer P., Abdulateef Y., Sullivan S., Walker J.F., Streeter D, & Burns J.M. (2023). First-Line Immunotherapy with Check-Point Inhibitors: Prospective Assessment of Cognitive Function. Cancers, 15(5), 1615.

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Nasal Biomarkers for Viral Respiratory Illness

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Grade 42 Whatman paper cut into similar-sized strips was placed into each nostril of the children for 2 minutes on each side. Samples were stored at −80°C until batched analysis could be performed. Filter papers were eluted using previously described methods.^{20 (link)} Luminex assays (MilliporeSigma, Burlington, Massachusetts) for periostin (detection limit = 0.12 ng/mL), eotaxin (detection limit = 4 pg/mL), interleukin (IL)-8 (detection limit = 0.4 pg/mL), and IL-13 (detection limit = 1.3 pg/mL) were used to determine levels of these cytokines. Comparisons of nasal inflammatory mediators and acute symptoms in VRAE and no virus subjects were performed.

Neumann N.F., Gyürek L.L., Gammie L., Finch G.R, & Belosevic M. (2000). Comparison of Animal Infectivity and Nucleic Acid Staining for Assessment of Cryptosporidium parvum Viability in Water. Applied and Environmental Microbiology, 66(1), 406-412.

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Cytokine Profiling of Activated T Cells

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The sorted cell populations were stimulated in 96-well plates with anti-CD3 (plate-bound) and anti-CD28 (soluble) antibodies or were left unstimulated for 4 days. Afterwards, cell supernatants were analyzed by Luminex assays (Millipore) for the following cytokines: Eotaxin, G-CSF, GM-CSF, IFN-α2, IL-10, IL-12p70, IFN-γ, IL-13, IL-17α, IL-1RA, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7,IL-8, IP10, MCP1, TNF-α, TNF-β, VEGF.

Viganò S., Negrón J.J., Tse S., Chowdhury F.Z., Lichterfeld M, & Yu X.G. (2017). HLA-G+ HIV-1-specific CD8 T cells are associated with HIV-1 immune control. AIDS (London, England), 31(2), 207-212.

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Cytokine and Receptor Profile Assessment

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Cytokines and their receptors [TNFα, soluble TNF receptor 1 (sTNFR1), soluble TNF Receptor 2 (sTNFR2), IFNy, IL-1β, interleukin-2 (IL-2), soluble interleukin-2 receptor (sIL-2R), IL-6, soluble interleukin-6 receptor (sIL-6R), interleukin-8 (IL-8), interleukin-10 (IL-10)] will be assessed via standard Luminex assays (high sensitivity assays from Millipore). We will measure samples in duplicate.

Loh K.P., Sanapala C., Janelsins M., Klepin H.D., Schnall R., Culakova E., Sohn M.B., Vertino P., Susiarjo M., Jensen-Battaglia M., Becker M.W., Liesveld J., Mendler J.H., Huselton E., Lin P.J, & Mustian K. (2021). Protocol for a pilot randomized controlled trial of a mobile health exercise intervention for older patients with myeloid neoplasms (GO-EXCAP 2). Journal of geriatric oncology, 13(4), 545-553.

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Cytokine Profiling in Statin-Treated Patients

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The levels of 24 cytokines/chemokines associated with innate (GM-CSF, IFNα, IL-10, IL-1β, IL-6, IL-8, TNFα, CCL2, CCL3, Th1 (IFNγ, IL-12P40, IL-12P70, CXCL10, CXCL9, CCL19, Th17 (IL-17F, IL-17A, IL-22, IL-21, IL-23, IL-25, IL-27), and B cell immune response (CXCL12, CXCL13) were assessed in patient CSF and serum samples using the bead-based Luminex assays (Millipore). To minimize inter-assay variation, all measurements were performed on the same day in one complete experiment. CSF and serum for cytokine determinations were available for 13 patients who were receiving statins and 66 patients who were not treated with statins.

Bogovič P., Lusa L., Stupica D., Rojko T., Korva M., Avšič-Županc T., Strle K., Wormser G.P, & Strle F. (2018). Impact of pre-existing treatment with statins on the course and outcome of tick-borne encephalitis. PLoS ONE, 13(10), e0204773.

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Cytokine Profiling in Co-Culture

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IL-2, IL-6, IL-10, IL-12p40, TNF-α, and IFN-γ levels were quantified in the 24 h co-culture supernatants using Luminex assays (Millipore, Billerica, MA, USA).

Moreno-Fernandez M.E., Joedicke J.J, & Chougnet C.A. (2014). Regulatory T Cells Diminish HIV Infection in Dendritic Cells – Conventional CD4+ T Cell Clusters. Frontiers in Immunology, 5, 199.

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Profiling Soluble Immunological Molecules

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Supernatants from cell cultures were collected after 72 h. A panel of 40 soluble immunological molecules were tested, including: EGF, FGF, EoTX, TGF, G-CSF, FRAK, IFNa2, IFNg, GRO, IL-10, MCP-3, IL-12, MDC, IL-12 p70, IL-13, IL-15, sCD40L, IL-17, IL-1RA, IL-1a, IL-9, IL1-B, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IP-10, MCP-1, MIP-1a, MIP-1B, TNFa, TNFB, VEGF, IL-22, MCP-2. Detection of those analytes was conducted using Luminex assays (Merck, Darmstadt, Germany). Assays were performed according to the manufacturer’s instructions. Data were collected and analyzed using Luminex software (Bio-Plex Manager Software, Standard Edition). A five-parameter regression formula was used to calculate the sample concentration from the standard curves.

Sanchez C., Jaramillo-Valverde L., Capristano S., Solis G., Soto A., Valdivia-Silva J., Poterico J.A, & Guio H. (2023). Antigen-Induced IL-1RA Production Discriminates Active and Latent Tuberculosis Infection. Microorganisms, 11(6), 1385.

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Intracellular Immune Markers in Serum and Synovial Fluid

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Cytotoxic and inflammatory mediators were determined by intracellular staining. Cells were fixed and permeabilized with Intracellular Fixation and Permeabilization Buffer Set (eBioscience), following the manufacturer’s instructions. To avoid non-specific binding, cells were incubated with 200 μl of permeabilization buffer containing 2% mouse serum (Sigma) for 20 min at RT. Without washing, 3 different antibody mixes were added. The first included Granzyme A-AF647 and IFNγ-PerCP/Cy5.5; the second, Granzyme B-AF647 and Perforin1-PerCP/Cy5.5, and the third, the respective isotype controls. Cells were incubated for 25 min at RT. After incubation, cells were washed and re-suspended in 200 μl of DPBS + 1% FBS for flow cytometry.
Protein levels of granzyme A, granzyme B, and perforin were determined in serum and SF (diluted 1:5 in PBS) using bead-based Luminex assays (EMD Millipore). All samples were assessed in one complete experiment to negate inter-assay variation.

Li T, & Rosazza J.P. (2000). Biocatalytic Synthesis of Vanillin. Applied and Environmental Microbiology, 66(2), 684-687.

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Comprehensive Cytokine and Hormone Profiling

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Cytokine/chemokine concentrations (VEGF-A, Leptin, IL-6, IL-1β, TNF-α, EGF, MCP-1, sFasL, and IL-17A) were measured in plasma using Luminex assays (Merck Millipore, Darmstadt, Germany). Cytokine/chemokine concentrations (IL-4, IL-5, IL-6, IL-10, IL-2, IFNγ, TNF-α, MCP-1, sFasL, and IL-17A) were measured in cell supernatant from phorbol 12-myristate 13-acetate (PMA) /ionomycin stimulated splenocytes. Insulin and adiponektin levels were measured in plasma with ELISA kits from Mercodia and RnD, respectively. All assays were performed according to manufactures instructions and values below detection limit and non-detectable values were set to zero in all analyses.

Rattik S., Hultman K., Rauch U., Söderberg I., Sundius L., Ljungcrantz I., Hultgårdh-Nilsson A., Wigren M., Björkbacka H., Fredrikson G.N, & Nilsson J. (2015). IL-22 affects smooth muscle cell phenotype and plaque formation in apolipoprotein E knockout mice. Atherosclerosis, 242(2).

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