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Luminex multiplex assay

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The Luminex multiplex assay is a bead-based technology that allows simultaneous detection and quantification of multiple analytes in a single sample. It utilizes color-coded magnetic beads coated with specific capture antibodies or ligands to target and measure various biomolecules, such as proteins, cytokines, or nucleic acids, in a high-throughput manner.

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Lab products found in correlation

Bio plex manager, by Bio-Rad (2 mentions) Bio plex 200, by Bio-Rad (2 mentions) Cd11b m1 70, by Thermo Fisher Scientific (1 mentions) Cd45 30 f11, by BD (1 mentions) Ly6g 1a8, by BioLegend (1 mentions) Cd16 32 93, by Thermo Fisher Scientific (1 mentions) Ly6c hk1, by Thermo Fisher Scientific (1 mentions) Lsrfortessa, by BD (1 mentions) Ab260058, by Abcam (1 mentions)

15 protocols using luminex multiplex assay

1

Luminex-based Multiplex Assays for Signaling

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Samples were run using dedicated kits (Millipore) on a Luminex MAGPIX machine. The effects of JQ1 on MYC levels were assessed with a Luminex multiplex assay (#48–617MAG, Millipore) according to the manufacturer’s instructions. To assess the effects of JQ1 on phosphorylation levels of PI3K pathway components (PTEN S380, AKT S473, GSK3a S21, GSK3β S9, TSC2 S939, mTOR S2448, and P70S6K T412), protein lysates were examined with a Luminex multiplex assay (#48–611MAG Millipore), according to the manufacturer’s instructions.
Each sample was run in triplicate, using 20 μg of total protein per sample, using the Luminex MAGPIX machine. Results were normalized to beta-tubulin expression levels, and beta-tubulin magnetic beads, and antibody was purchased from Millipore (#46–713MAG).
Derenzini E., Mondello P., Erazo T., Portelinha A., Liu Y., Scallion M., Asgari Z., Philip J., Hilden P., Valli D., Rossi A., Djaballah H., Ouerfelli O., de Stanchina E., Seshan V.E., Hendrickson R.C, & Younes A. (2018). BET Inhibition-Induced GSK3β Feedback Enhances Lymphoma Vulnerability to PI3K Inhibitors. Cell reports, 24(8), 2155-2166.
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2

Cytokine and Angiogenic Profiles in CM

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Cytokine, chemokine, and angiogenic growth factor levels were tested uNK-CM, PEx-CM, uNK/PEx-dir-CM and uNK/PEx-indir-CM all with or without 1,25(OH)2D (0, 1, and 10 nM) or 25OHD (0, 10, and 100 nM) (n = 5 each group) by Luminex multiplex assay according to the manufacturer’s instructions (Millipore). The analytes tested were EGF, IFN-γ, IL-1RA, IL-1β, IL-4, IL-6, MDC, PDGF-AA, PDGF-BB, VEGF-A, FGF-2, MCP-1, MCP-3, MIP-1α, MIP-1β, RANTES, TNF-α, PLGF, IP-10, G-CSF, GM-CSF, GRO-α, and Ang-2.
Zhang J.Y., Wu P., Chen D., Ning F., Lu Q., Qiu X., Hewison M., Tamblyn J.A., Kilby M.D, & Lash G.E. (2020). Vitamin D Promotes Trophoblast Cell Induced Separation of Vascular Smooth Muscle Cells in Vascular Remodeling via Induction of G-CSF. Frontiers in Cell and Developmental Biology, 8, 601043.
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3

Measuring Serum Triglycerides and Insulin

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Serum triglycerides were measured using a FUJI DRI-CHEM 3000 (Fuji Photo Film Co., Tokyo, Japan). Serum insulin concentrations were quantified using a Luminex multiplex assay (cat#: MENDO-75K-05, Millipore Corporation, Bedford, MA). The homeostasis model assessment for insulin resistance (HOMA-IR = fasting blood glucose [mM] × fasting insulin [μU/mL]/22.5) and the homeostasis model assessment for beta-cell function (HOMA-B = 20 × fasting insulin [μU/mL]/fasting blood glucose [mM] − 3.5) were then assessed [23 (link)].
Chang C.C., Yuan W., Lin Y.L., Liu R.S., Juan Y.C., Sun W.H., Tsay H.J., Huang H.C., Lee Y.C, & Liu H.K. (2016). Evaluation of the In Vivo Therapeutic Effects of Radix Paeoniae Rubra Ethanol Extract with the Hypoglycemic Activities Measured from Multiple Cell-Based Assays. Evidence-based Complementary and Alternative Medicine : eCAM, 2016, 3262790.
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4

Multiplex Assay for Gut Hormones

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Plasma samples, standards and controls were tested in duplicate using an active ghrelin ELISA, and active amylin and gastric inhibitory polypeptide (GIP) were measured by Luminex multiplex assay (Millipore, Billerica, MA). For active ghrelin, samples were tested undiluted and plates were prepared according to protocol and quantified using a 6-point standard curve ranging from 172 to 5500 ng/mL. Plates were read using a Molecular Devices Plate reader and Softmax Pro data analysis software. A 5-PL curve fit was used. Data analysis was performed using Softmax Pro 5.0. For amylin and GIP, samples, standards and controls were tested in duplicate and the assay was prepared according to protocol using a 7 point standard curve. Plates were read using the Bioplex 200 with Bioplex Manager (BioRad, Hercules, CA).
McFarlane S.I., Mielke M.M., Uglialoro A., Keating S.M., Holman S., Minkoff H., Crystal H.A, & Gustafson D.R. (2017). Ghrelin, Amylin, Gastric Inhibitory Peptide and Cognition in Middle-Aged HIV-Infected and Uninfected Women: The Women’s Interagency HIV Study. Journal of neurology & neurophysiology, 8(1), 413.
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5

Multiplex Immune Marker Analysis

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Ten immune markers were measured in duplicate in all samples (n=260) by a Luminex multiplex assay from Millipore (USA): MCP-3, MIP-1α, MIP-1β, VEGF, FGF-2, fractalkine, TGF-α, IL-13, TNF-α, and IL-10. Samples from matched cases and controls were included in random order in the same analytical batch. Laboratory personnel were blinded concerning case-control status and chronological order of samples. All analyses were performed according to the manufacturer’s protocol (Online Supplementary Methods).
M-proteins were assessed in samples from all future myeloma cases except four (due to insufficient sample volumes) by protein electrophoresis, immunofixation electrophoresis, and FLC assays (Online Supplementary Methods).
Späth F., Wibom C., Krop E.J., Santamaria A.I., Johansson A.S., Bergdahl I.A., Hultdin J., Vermeulen R, & Melin B. (2019). Immune marker changes and risk of multiple myeloma: a nested case-control study using repeated pre-diagnostic blood samples. Haematologica, 104(12), 2456-2464.
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6

Measuring IRBP-Responsive T Cell Cytokines

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To measure cytokine production of IRBP-responsive T cells, single cell suspensions were prepared as described above and were stimulated for 48h with 20 μg/ml whole IRBP protein; after which the supernatants were collected for analysis of cytokine levels. Concentration of cytokines (IFNγ, IL-17A, IL-10, IL-12p40, IP-10, KC, TNFα, IL-6, and IL-2) was measured using a Luminex® multiplex assay (Millipore, Billerica, MA, USA) on Model L100IS (Luminex, Austin, TX, USA) and analyzed with BeadView™ (Upstate Cell Signaling Solutions, Lake Placid, NY, USA) as described (25 (link)).
Lee E.J., Brown B.R., Vance E.E., Snow P.E., Silver P.B., Heinrichs D., Lin X., Iwakura Y., Wells C.A., Caspi R.R, & Rosenzweig H.L. (2016). Mincle activation and the Syk/Card9 signaling axis are central to development of autoimmune disease of the eye. Journal of immunology (Baltimore, Md. : 1950), 196(7), 3148-3158.
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7

Cytokine Analysis in Acute Sepsis

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Convenience blood samples were obtained on Day 1 and/or Day 4 of SAB where available for cytokine analysis. Samples were centrifuged and stored at − 80 °C until further analyses. Cytokine concentrations (IFNγ, IL-1β, IL-6, IL-8, IL-10, IL-17, TNF) were measured in duplicates using Luminex multiplex assay (Millipore, Billerica, MA) per manufacturer’s instructions.
Tan K., Minejima E., Lou M., Mack W.J., Nieberg P, & Wong-Beringer A. (2021). Cytokine measurements add value to clinical variables in predicting outcomes for Staphylococcus aureus bacteremia. BMC Infectious Diseases, 21, 317.
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8

Cytokine Profiling in Sepsis

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Plasma or serum samples were collected from specimens drawn for routine labs once physician-ordered tests were completed. Samples were collected at onset of SAB (within 24h of the index positive blood culture) and at 72h after starting effective antibiotic therapy and stored at -80°C until analysis. Cytokine concentrations were determined by Luminex® multiplex assay according to manufacturer’s instructions (Millipore, Billerica, MA) for pro-inflammatory cytokines: TNF, IL-6, IL-8, IL-17A, and anti-inflammatory cytokine: IL-10. Individual ratio of IL-10 to each pro-inflammatory mediator was calculated for each patient at both time points. Samples were allowed to thaw to room temperature only once before testing. All assays were performed in duplicate.
Minejima E., Bensman J., She R.C., Mack W.J., Tran M.T., Ny P., Lou M., Yamaki J., Nieberg P., Ho J, & Wong-Beringer A. (2016). A dysregulated balance of pro- and anti-inflammatory host cytokine response early during therapy predicts persistence and mortality in Staphylococcus aureus bacteremia. Critical care medicine, 44(4), 671-679.
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9

Multiplex Assay for Gut Hormones

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Plasma samples, standards and controls were tested in duplicate using an active ghrelin ELISA, and active amylin and gastric inhibitory polypeptide (GIP) were measured by Luminex multiplex assay (Millipore, Billerica, MA). For active ghrelin, samples were tested undiluted and plates were prepared according to protocol and quantified using a 6-point standard curve ranging from 172 to 5500 ng/mL. Plates were read using a Molecular Devices Plate reader and Softmax Pro data analysis software. A 5-PL curve fit was used. Data analysis was performed using Softmax Pro 5.0. For amylin and GIP, samples, standards and controls were tested in duplicate and the assay was prepared according to protocol using a 7 point standard curve. Plates were read using the Bioplex 200 with Bioplex Manager (BioRad, Hercules, CA).
McFarlane S.I., Mielke M.M., Uglialoro A., Keating S.M., Holman S., Minkoff H., Crystal H.A, & Gustafson D.R. (2017). Ghrelin, Amylin, Gastric Inhibitory Peptide and Cognition in Middle-Aged HIV-Infected and Uninfected Women: The Women’s Interagency HIV Study. Journal of neurology & neurophysiology, 8(1), 413.
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10

Multiplex Cytokine and Complement Profiling in Murine TRALI

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Joint fluid and sera were evaluated for cytokine concentrations by a Luminex-Multiplex assay (Millipore) according to the manufacturer's directions. Flow cytometric analysis of blood and joint digests was performed as described (44) . Briefly, single-cell suspensions were stained with the following: Ly6C (HK1.4, eBioscience), Ly6G (1A8, BioLegend), CD11b (M1/70, eBioscience), CD16/32 (93, eBioscience), CD64 (X54-5/7.1.1; BD Pharmingen), and CD45 (30-F11, BD Pharmingen). The cells were fixed and acquired on a BD LSRFortessa and analyzed with FlowJo software. Joint washes and sera were evaluated for C1q, C5a (R&D Systems), and C3 (GENWAY) enzyme-linked immunosorbent assays. The ability of Ter119 and deglycosylated Ter119 to passively sensitize erythrocytes in vivo on days 1 and 3 after antibody injection was assessed ex vivo by flow cytometry using an anti-rat-PE-conjugated antibody.
Transfusion-related acute lung injury TRALI was induced as described (77) . Briefly, SCID mice were injected with 40 of g Ter119 24 hours before injection of 50 g of the TRALI-inducing antibody 34-1-2s. Rectal temperatures were recorded every 30 min for 2 hours, and mice were then euthanized to determine lung W/D weight ratios (77) . Pulmonary neutrophils were enumerated as previously described (77) .
Crow A.R., Kapur R., Koernig S., Campbell I.K., Jen C.C., Mott P.J., Marjoram D., Khan R., Kim M., Brasseit J., Cruz-Leal Y., Amash A., Kahlon S., Yougbare I., Ni H., Zuercher A.W., Käsermann F., Semple J.W, & Lazarus A.H. (2019). Treating murine inflammatory diseases with an anti-erythrocyte antibody. Science translational medicine, 11(506).
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