Fluorokine multianalyte profiling kit

Manufactured by R&D Systems

Sourced in United States

The Fluorokine multianalyte profiling kit is a laboratory equipment product designed for the simultaneous quantitative measurement of multiple analytes in a single sample. The kit utilizes fluorescence-based detection technology to enable the analysis of various proteins, cytokines, or other biomolecules in a high-throughput manner.

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Lab products found in correlation

Luminex 200 platform, by Bio-Rad (2 mentions) Bio plex 200 system, by Bio-Rad (1 mentions) Bio plex 200 platform, by Bio-Rad (1 mentions) Milliplex non human primate cytokine chemokine kit, by Merck Group (1 mentions) Luminex xmap multiplexed bead system, by Merck Group (1 mentions) Luminex platform, by Bio-Rad (1 mentions) Luminex 200 analyzer, by R&D Systems (1 mentions) Ensight, by PerkinElmer (1 mentions) Igf 2, by Mediagnost (1 mentions) Lumipulse g kl 6, by Fujirebio (1 mentions) Bioplex 2200 analyzer, by Bio-Rad (1 mentions)

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Fluorokine-1. Multianalyte Profiling Kits (2 citations)

13 protocols using fluorokine multianalyte profiling kit

Cytokine Profiling of Skin Samples

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At indicated time points, skin samples (4 mm) were collected, minced, and placed into Cell Lysate Buffer (RayBiotech; Norcross, GA) supplemented with protease inhibitors. Lysates were diluted 1:2 and cytokine concentrations were measured in duplicate utilizing Luminex technology with the Fluorokine® Multianalyte Profiling kit according to manufacturer’s instructions (R&D systems; Minneapolis, MN). Samples were read on a Bio-Plex 200 system (BioRad; Hercules, CA) using the Bioplex 6.1 software.

Diaz-Perez J.A., Killeen M.E., Yang Y., Carey C.D., Falo LD J.r, & Mathers A.R. (2018). Extracellular ATP and IL-23 form a local inflammatory circuit leading to the development of a neutrophil-dependent psoriasiform dermatitis. The Journal of investigative dermatology, 138(12), 2595-2605.

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Multiplex Profiling of Neuroinflammatory Mediators

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Adhesion molecules, cytokines and chemokines were analysed by Fluorokine multianalyte profiling kit according to the manufacturer’s protocol (R&D Systems, Minneapolis, Minnesota) on the Bio-Plex 200 platform (Bio-Rad, Hercules, California). The minimum detection limit for the ICAM-1 and p-selectin were 52.7 pg/ml and 2.6 pg/ml, respectively. The minimum detection limit for the cytokines and chemokines were CCL-2/MCP-1 134 pg/ml, CCL-3/MIP-1α 0.452 pg/ml, CCL-4/ MIP-1β 77.4 pg/ml, CCL-5/ RANTES 19.1 pg/ml, CCL-7/ MCP-3 1.69 pg/ml, CCL-8/ MCP-2 0.283 pg/ml, CCL-11/Eotaxin 1.46 pg/m, CCL-12/ MCP-5 0.613 pg/ml, CCL-19/ MIP-3β 2.28 pg/ml, CCL-20/ MIP-3α 3.95 pg/ml, CCL-22/ MDC 0.965 pg/ml, CXCL-1/ KC 32.9 pg/ml, CXCL-2/ MIP-2 1.97 pg/ml, CXCL-10/ IP-10 6.85 pg/ml, CXCL-13/ BLC 19.3 pg/ml, IL-1α 8.17 pg/ml, IL-1β 41.8 pg/ml, IL-6 2.30 pg/ml, IL-12 p70 12.8 pg/ml, IL-17A 7.08 pg/ml, IL-27 1.84 pg/ml, LIX 2.02 pg/ml, TNF-α 1.47 pg/ml, IFN-γ 1.85 pg/ml. Brain homogenates were assayed at neat for all analytes and results were normalised to their total protein concentrations (Bio-Rad, Hercules, California).

Poh X.Y., Hong J.M., Bai C., Miow Q.H., Thong P.M., Wang Y., Rajarethinam R., Ding C.S, & Ong C.W. (2022). Nos2−/− mice infected with M. tuberculosis develop neurobehavioral changes and immunopathology mimicking human central nervous system tuberculosis. Journal of Neuroinflammation, 19, 21.

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Multiplex Cytokine Profiling Assay

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Measurements of IL-6, IL-8 and IL-10 were performed using a Fluorokine Multi-analyte Profiling Kit (catalog number base kit: LUH000; IL-6: LUH206; IL-8: LUH208 and IL-10: LUH217) from R&D Systems, Minneapolis, MN, USA following the manufacturer’s instructions. Lipopolysaccharide (LPS) at 10µg/ml was employed as positive control. Negative controls used for the cytotoxicity experiments remained the same. Cytokine concentrations were determined using the dual laser flow analyzer Luminex 200 (Luminex Corp, Austin, TX, USA). Standard curve for each cytokine was plotted employing a 5-paremeter logistic fit (5-PL). This has been previously reported by our laboratory (Fuentes-Mattei et al., 2010 ; Rodriguez-Cotto et al., 2013 ).

Rodríguez-Cotto R.I., Ortiz-Martínez M.G., Rivera-Ramírez E., Mateus V.L., Amaral B.S., Jiménez-Vélez B.D, & Gioda A. (2014). Particle Pollution in Rio de Janeiro, Brazil: Increase and Decrease of Pro-inflammatory Cytokines IL-6 and IL-8 in Human Lung Cells. Environmental pollution (Barking, Essex : 1987), 194, 112-120.

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Cytokine Profiling in Cell Cultures

Cited 1 time

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Cytokine analyses (IL-6, IL-8 and IL-10) were performed using a Fluorokine Multi-analyte Profiling Kit from R & D Systems, (Minneapolis, MN, USA) according to the manufacturer’s instructions. Lipopolysacharide (LPS), a positive control was used at a final concentration of 10 μg mL^-1. The same controls used for the cytotoxicity experiments were employed. Cytokine concentrations were determined using the dual laser flow analyzer Luminex 200 (Luminex Corp, Austin, TX, USA). Standard curves for each cytokine were plotted employing a 5-paremeter logistic fit (5-PL). This procedure has been previously used and reported by our laboratory (Rodríguez-Cotto et al., 2014 (link)).

Gioda A., Rodríguez-Cotto R.I., Amaral B.S., Encarnación-Medina J., Ortiz-Martínez M.G, & Jiménez-Vélez B.D. (2016). Biodiesel from Soybean Promotes Cell Proliferation in Vitro. Toxicology in vitro : an international journal published in association with BIBRA, 34, 283-288.

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Cytokine and Chemokine Profiling of HIV-1 Infection

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Cytokines and chemokines concentration (IL2, IFNγ, MCP-1, MIP1α, MIP1β and RANTES) was assessed in supernatants of uninfected and HIV-1 infected cells cultures using multiplex sandwich immunoassays (Fluorokine Multi Analyte Profiling Kit; R&D Systems, Minneapolis, MN, USA) according to manufacturer’s protocol. Supernatants were obtained after 7 days of infection.

Trabattoni D., Gnudi F., Ibba S.V., Saulle I., Agostini S., Masetti M., Biasin M., Rossignol J.F, & Clerici M. (2016). Thiazolides Elicit Anti-Viral Innate Immunity and Reduce HIV Replication. Scientific Reports, 6, 27148.

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Multiplexed Cytokine and Chemokine Quantification

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Cytokines (TNF-α and IL-10) and the βeta (β)-chemokine-MIP-1β were measured simultaneously with either the Milliplex Non-human Primate Cytokine/Chemokine kit (Millipore) or the Fluorokine Multianalyte Profiling kit (R&D) using the Luminex xMAP multiplexed bead system (Millipore), according to the manufacturer's instructions. Results obtained from the Luminex xMAP system were analysed automatically by the Luminex xPONENT software programme (Millipore) using a standard curve derived from recombinant cytokine and chemokine standards.

Pegu A., Asokan M., Wu L., Wang K., Hataye J., Casazza J.P., Guo X., Shi W., Georgiev I., Zhou T., Chen X., O'Dell S., Todd J.P., Kwong P.D., Rao S.S., Yang Z.Y., Koup R.A., Mascola J.R, & Nabel G.J. (2015). Activation and lysis of human CD4 cells latently infected with HIV-1. Nature Communications, 6, 8447.

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Fluorokine Multianalyte MMP-8 and -9 Assay

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MMP-8 and -9 concentrations were analyzed by Fluorokine multianalyte profiling kit according to the manufacturer’s protocol (R&D Systems) on the Luminex 200 platform (Bio-Rad). The minimum level of detection for MMP-8 and -9 was 110 pg/ml and 65 pg/ml respectively.

Ong C.W., Fox K., Ettorre A., Elkington P.T, & Friedland J.S. (2018). Hypoxia increases neutrophil-driven matrix destruction after exposure to Mycobacterium tuberculosis. Scientific Reports, 8, 11475.

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MMP-8/-9 Concentrations by Luminex

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MMP-8 and -9 concentrations were analyzed by Fluorokine multianalyte profiling kit according to the manufacturer’s protocol (R&D Systems) on the Luminex platform (Bio-Rad). The minimum level of detection for MMP-8 and -9 was 110 pg/ml and 65 pg/ml respectively. Cytokine concentrations were analyzed using a human 30-plex panel (Invitrogen).

Ong C.W., Elkington P.T., Brilha S., Ugarte-Gil C., Tome-Esteban M.T., Tezera L.B., Pabisiak P.J., Moores R.C., Sathyamoorthy T., Patel V., Gilman R.H., Porter J.C, & Friedland J.S. (2015). Neutrophil-Derived MMP-8 Drives AMPK-Dependent Matrix Destruction in Human Pulmonary Tuberculosis. PLoS Pathogens, 11(5), e1004917.

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Plasma Inflammation and Lipid Peroxidation

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The inflammation level was evaluated on patients’ plasma samples (blood samples centrifuged at 4000 rpm at 4 °C for 20 min). In particular, eight cytokines/chemokines (IL-1β, IL-4, IL-6, IL-12p70, IL-18, MCP-1, TNF-α, and INF-γ) were measured using the ELISA (enzyme-linked immunosorbent assays) immunological test. We used the Fluorokine^® Multianalyte Profiling kit (R & D Systems, Minneapolis, MN, USA) through the Luminex^® 200 analyzer, according to the manufacturing procedures.
To assess lipid peroxidation level in plasma samples, a Parameter™ TBARS (thiobarbituric-acid-reactive substances) assay was used (R & D Systems, Minneapolis, MN, USA), according to the manufacturing procedures. Briefly, in the presence of heat and acid, malondialdehyde (MDA) reacts with TBA to produce a colored end-product that absorbs light at 530–540 nm. The intensity of the color at 532 nm corresponds to the level of lipid peroxidation in the sample, measured through a multimode microplate reader, EnSight™ (PerkinElmer, Beaconsfield, UK).

Pivari F., Mingione A., Piazzini G., Ceccarani C., Ottaviano E., Brasacchio C., Dei Cas M., Vischi M., Cozzolino M.G., Fogagnolo P., Riva A., Petrangolini G., Barrea L., Di Renzo L., Borghi E., Signorelli P., Paroni R, & Soldati L. (2022). Curcumin Supplementation (Meriva®) Modulates Inflammation, Lipid Peroxidation and Gut Microbiota Composition in Chronic Kidney Disease. Nutrients, 14(1), 231.

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Quantifying MMP-8 and MMP-9 via Fluorokine Kit

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MMP-8 and -9 concentrations were analyzed by Fluorokine MultiAnalyte Profiling Kit according to manufacturer’s protocol (R&D Systems) on the Luminex 200 platform (Bio-Rad). The minimum level of detection for MMP-8 and -9 was 110 and 65 pg/ml, respectively.

Ong C.W., Pabisiak P.J., Brilha S., Singh P., Roncaroli F., Elkington P.T, & Friedland J.S. (2017). Complex regulation of neutrophil-derived MMP-9 secretion in central nervous system tuberculosis. Journal of Neuroinflammation, 14, 31.

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