Luminex

Manufactured by Eve Technologies

Sourced in Canada

The Luminex lab equipment is a multi-analyte profiling system that uses proprietary xMAP technology to simultaneously measure multiple analytes in a single sample. It enables rapid and accurate detection and quantification of a variety of biomolecules, including proteins, nucleic acids, and small molecules, in a high-throughput manner.

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Lab products found in correlation

Duoset elisa kit, by R&D Systems (1 mentions) Interleukin 4 (il 4), by R&D Systems (1 mentions) Stereotaxic device, by Kopf Instruments (1 mentions) Atp determination kit, by Thermo Fisher Scientific (1 mentions) Arl67156, by Merck Group (1 mentions) Mouse lipocalin 2 duoset elisa kit, by R&D Systems (1 mentions) Ldh colorimetric assay kit, by Abcam (1 mentions) Serum separator tube, by BD (1 mentions) Quickplex sq 120, by MSD (1 mentions)

10 protocols using luminex

Cytokine and Chemokine Profiling in Liver

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Hepatic levels of the cytokines TNFα and IL-6, and neutrophil-relevant chemokines CXCL1 and CXCL2, were measured in liver homogenates by Luminex® (Eve Technologies Corporation, Calgary, Canada) (28 (link)). Liver homogenate protein concentrations were quantified using a BCA Protein Assay kit (Pierce, USA). Results expressed as pg/mg protein.

Almishri W., Shaheen A.A., Sharkey K.A, & Swain M.G. (2019). The Antidepressant Mirtazapine Inhibits Hepatic Innate Immune Networks to Attenuate Immune-Mediated Liver Injury in Mice. Frontiers in Immunology, 10, 803.

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Quantifying Vaginal Cytokine Levels

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Vaginal lavages were diluted in PBS and analysed for cytokine levels. Lavages subjected to a 32-plex Luminex (Eve Technologies) including IL-4, IL-5, IL-6, IL-9, IL-13, IFN-γ, TNF-α, and IL-1β. IFN-γ and IL-6 were independently quantified in vaginal lavages using R&D DuoSet ELISA kits. IFN-α was independently quantified using the R&D ELISA kit.

Lee A.J., Feng E., Chew M.V., Balint E., Poznanski S.M., Giles E., Zhang A., Marzok A., Revill S.D., Vahedi F., Dubey A., Ayaub E., Jimenez-Saiz R., McGrath J.J., Ritchie T.M., Jordana M., Jonigk D.D., Ackermann M., Ask K., Miller M., Richards C.D, & Ashkar A.A. (2022). Type I interferon regulates proteolysis by macrophages to prevent immunopathology following viral infection. PLoS Pathogens, 18(5), e1010471.

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Polarization and Cytokine Analysis of BMDM

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BMDM were seeded in 6-well plates at 2×10⁶ cells/well and polarized for 48 hours with the addition of 50 ng/ml LPS (List Biological Labs) and 50 ng/ml IFNγ (Cedarlane) for M1, or 40 ng/ml IL-4 (R&D Systems) for M2 at 37°C in a 5% CO₂ humidified atmosphere. Cells were re-plated in triplicate in 96-well plates, 8×10⁵ cells/well, in media containing 20 µg/ml DMXAA or DMSO control for 24 hours. Supernatants were subjected to a mouse 32-plex cytokine/chemokine discovery array (Luminex) (EVE Technologies, Calgary, Alberta).

Downey C.M., Aghaei M., Schwendener R.A, & Jirik F.R. (2014). DMXAA Causes Tumor Site-Specific Vascular Disruption in Murine Non-Small Cell Lung Cancer, and like the Endogenous Non-Canonical Cyclic Dinucleotide STING Agonist, 2′3′-cGAMP, Induces M2 Macrophage Repolarization. PLoS ONE, 9(6), e99988.

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Measuring ATP and IL-1β in Rat CSF

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CSF was obtained from the rats, as previously reported by Pegg et al. (47 (link)). Briefly, rats were anesthetized with 2% (v/v) isoflurane and placed in a stereotaxic device (Kopf Instruments). The head and neck were shaved, and a midline incision was made to expose the cisterna magna and the area blotted dry of blood. Using pulled capillary tubing, a tube was inserted through the dura mater at a parallel angle to collect CSF. CSF volume ranged from 75 to 150 μl and was stored in a 0.5-ml tube at −80°C with 1 μM ARL67156 (Sigma) to minimize ATP degradation. Samples contaminated with blood were not included in this study. ATP levels were detected using an ATP Determination kit (Life Technologies). The samples were combined with firefly luciferase and its substrate d-luciferin. Using bioluminescence, the ATP concentration was detected using a FilterMax F5 place reader at 27°C. The relative concentration of IL-1β in rat CSF was measured using antibody-based multiplex immunoassays (Luminex, Eve Technologies).

Mousseau M., Burma N.E., Lee K.Y., Leduc-Pessah H., Kwok C.H., Reid A.R., O’Brien M., Sagalajev B., Stratton J.A., Patrick N., Stemkowski P.L., Biernaskie J., Zamponi G.W., Salo P., McDougall J.J., Prescott S.A., Matyas J.R, & Trang T. (2018). Microglial pannexin-1 channel activation is a spinal determinant of joint pain. Science Advances, 4(8), eaas9846.

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Cytokine-Modulated Antigen-Specific T-Cell Response

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To assess the influence of cytokine co-administration, recombinant GM-CSF, IL-21 or their combination were administered to animals along with OVA-pulsed MSC-IPr injection. Cytokines were administered every 48 h (three injections at week 1) and the same cycle was repeated at week 3 for a total of 6 injections. Sera were collected weekly, whereas cytokine analysis was conducted at the end of the experiment (week 16). The obtained CD4 T-cells were then co-cultured for 72 h with MSC-IPr previously pulsed with ISQAVHAAHAEINEAGR (1 μg/mL). Supernatants were then collected and analyzed by Luminex at Eve Technologies (Calgary, AB, CA) to evaluate the cytokine and chemokine secretion profile of the responding CD4 T-cells.

Bikorimana J.P., Abusarah J., Salame N., El-Hachem N., Shammaa R, & Rafei M. (2022). Humoral Immunity to Allogeneic Immunoproteasome-Expressing Mesenchymal Stromal Cells Requires Efferocytosis by Endogenous Phagocytes. Cells, 11(4), 596.

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Serum Biomarker Profiling in Mice

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All assays were performed using hemolysis-free sera, which were collected by centrifugation of blood in serum separator tubes (BD Biosciences, USA). Serum LDH levels were determined by colorimetric LDH assay kit (Abcam, ON, Canada) as per manufacturer’s protocol. Serum lipocalin 2 was detected and quantified using mouse lipocalin 2 duoset ELISA kit (R&D systems; Catalog No. DY1857-05). Levels of anti-flagellin and LPS antibodies in serum was detected by ELISA^{58 (link)}. Serum cytokine levels were measured using luminex (Eve technologies, University of Calgary, Calgary). Total iron content was quantified using hemolysis-free sera and from saline-perfused liver samples from animals. Iron quantification kit (Sigma, USA) was used to determine total iron content following manufacturer’s protocol.

Kumar M., Leon Coria A., Cornick S., Petri B., Mayengbam S., Jijon H.B., Moreau F., Shearer J, & Chadee K. (2020). Increased intestinal permeability exacerbates sepsis through reduced hepatic SCD-1 activity and dysregulated iron recycling. Nature Communications, 11, 483.

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Multiplex Cytokine Profiling of Serum and Cell Culture Samples

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Serum samples or culture media were processed and subsequently frozen at −80 °C until use. Serum samples were analyzed either by luminescence (Luminex, Eve Technologies, Calgary, Canada) or electro chemiluminescence assays using the Meso-Scale Discovery (IFNγ or IL-27p28) or mouse custom Multiplex kit for 10 or 27 factors (Mesoscale Diagnostic, MSD). All samples were tested in duplicate and read on MSD QuickPlex SQ 120 imager/reader. All cytokine values were reported in units of pg/mL. See Supplementary Methods for further detail.

Valencia J.C., Erwin-Cohen R.A., Clavijo P.E., Allen C., Sanford M.E., Day C.P., Hess M.M., Johnson M., Yin J., Fenimore J.M., Bettencourt I.A., Tsuneyama K., Romero M.E., Klarmann K.D., Jiang P., Bae H.R., McVicar D.W., Merlino G., Edmondson E.F., Anandasabapathy N, & Young H.A. (2021). Myeloid derived suppressive cell expansion promotes melanoma growth and autoimmunity by inhibiting CD40/IL-27 regulation in macrophages. Cancer research, 81(23), 5977-5990.

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Murine Macrophage Cytokine Response

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Peritoneal exudate cells (PECs) from naïve male mice were collected by lavage with 8 ml/mouse RPMI-2% FBS and then plated on culture dishes with complete RPMI. After incubating overnight, the dishes were washed 4 times with DPBS to remove non-adherent cells and the adherent cells were incubated with DPBS-5 mM EDTA at 37°C for 7 min. The macrophages were then pipetted off the dish with complete RPMI. The purity of macrophages (CD11b^hi, F4/80^hi) was greater than 97% (data not shown). In 96-well plates, 1x10⁵/well macrophages were stimulated with 30 μg/ml histones and/or 0.1 μg/ml CpG1826 ODN for overnight. The culture supernatant was then harvested and assayed for cytokines by Luminex (Eve Technologies, Calgary, AB, Canada) to detect inflammatory mediators.

Lai J.J., Cruz F.M, & Rock K.L. (2019). Immune Sensing of Cell Death through Recognition of Histone Sequences by C-Type Lectin-Receptor-2d Causes Inflammation and Tissue Injury. Immunity, 52(1), 123-135.e6.

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Quantification of Inflammatory Mediators

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Levels of IL-1Ra in serum and single hind paws were assessed by ELISA (Quantikine-IL1Ra, R&D Systems). Levels of paw and serum inflammatory mediators were evaluated using Luminex (Mouse High Sensitivity T cell Discovery Array 18-Plex, Eve Technologies, Calgary, AB, Canada).

Choi Y.R., Collins K.H., Springer L.E., Pferdehirt L., Ross A.K., Wu C.L., Moutos F.T., Harasymowicz N.S., Brunger J.M., Pham C.T, & Guilak F. (2021). A genome-engineered bioartificial implant for autoregulated anticytokine drug delivery. Science Advances, 7(36), eabj1414.

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Cytokine and Chemokine Profiling of MSCs

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For cytokine and chemokine profiling, 15 cm culture petri dishes containing 80–90% confluent MSC-β5t or Ctl MSCs were grown in serum-free media for 24 h at 37°C. Collected supernatants were then analyzed using luminex by Eve Technologies (Calgary, AB, CA) or commercially-available chemokine arrays (RayBiotech) according to manufacturer’s instructions.

Bikorimana J.P., El-Hachem N., El-Kadiry A.E., Abusarah J., Salame N., Shammaa R, & Rafei M. (2021). Thymoproteasome-Expressing Mesenchymal Stromal Cells Confer Protective Anti-Tumor Immunity via Cross-Priming of Endogenous Dendritic Cells. Frontiers in Immunology, 11, 596303.

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