Luminex 200tm instrument

Manufactured by DiaSorin

Sourced in United States

The Luminex 200TM instrument is a multiplex assay platform that utilizes Luminex xMAP technology to perform simultaneous detection and quantification of multiple analytes in a single sample. The core function of the instrument is to facilitate high-throughput, multiplexed bioassays through the use of color-coded magnetic beads.

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Lab products found in correlation

Milliplex map human glycolysis pathway magnetic bead panel hgpmag 27k, by Merck Group (2 mentions) Luminex xmap technology, by DiaSorin (1 mentions) Multiplex assay technology, by DiaSorin (1 mentions) Elisa kit, by R&D Systems (1 mentions) Procartaplex multiplex kit, by Thermo Fisher Scientific (1 mentions) Xponent software, by DiaSorin (1 mentions) Elx405 washer, by Agilent Technologies (1 mentions)

6 protocols using luminex 200tm instrument

Simultaneous Quantification of Glycolytic Enzymes

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The MILLIPLEX^® MAP Human Glycolysis Pathway Magnetic Bead Panel (HGPMAG-27K, Millipore) was applied in 96-well plates for the simultaneous quantification of the following proteins in tissue lysates: G6PI/AMF (Glucose-6-phosphate isomerase/Autocrine motility factor), LDHA (L-lactate dehydrogenase A chain), LDHB (L-lactate dehydrogenase B chain), PKM2 (Pyruvate kinase isoform M2) and TKT (Transketolase), and HIF-1α (Hypoxia-inducible factor 1-α).
For the immunoassay procedures, 25μl of each dilute lysate sample in Assay Buffer (5μg total protein/well) and HeLa cells lysate (positive control) were added into wells in duplicate, according to the manufacturer's instructions. To each well, 25 μl of the Mixed Beads were added and the plate was incubated for 2 hours at room temperature. Human glycolysis pathway detection biotinylated antibodies were added for 1 hour; each captured a specific bead. After that, the reaction mixture was incubated for 30 minutes with Streptavidin-PE conjugate to complete the reaction on the surface of each microsphere. Finally, the MILLIPLEX^® MAP was analyzed by Luminex xMAP^® technology. The immunoassay on the surface of each fluorescent-coded magnetic bead, MagPlex-C microsphere, was identified and quantified based on fluorescent signals. The median fluorescence intensity (MFI) was read with the Luminex 200^TM instrument and measured with xPONENT^® software.

Lucarelli G., Galleggiante V., Rutigliano M., Sanguedolce F., Cagiano S., Bufo P., Lastilla G., Maiorano E., Ribatti D., Giglio A., Serino G., Vavallo A., Bettocchi C., Selvaggi F.P., Battaglia M, & Ditonno P. (2015). Metabolomic profile of glycolysis and the pentose phosphate pathway identifies the central role of glucose-6-phosphate dehydrogenase in clear cell-renal cell carcinoma. Oncotarget, 6(15), 13371-13386.

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Multiplex Assay for Measuring IL-6 Levels

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For IL‐6 analysis of cell and tissue extracts, samples were assessed using the multiplex assay technology by Luminex as described previously.²², ²³ The levels of IL‐6 were determined by standard curve analysis. The plate was read on a Luminex 200TM instrument (Luminex, Austin, TX). Data acquisition and analysis were conducted using StarStation software v2.3 (Applied Cytometry Systems, Dinnington, UK). In addition, cell culture supernatants were collected from MSCs, and SDF‐1 was measured using a commercially available ELISA kit (R&D, Minneapolis, MN) according to the manufacturer's instructions.²⁴

Kwon M., Ghanta S., Ng J., Castano A.P., Han J., Ith B., Lederer J.A., El‐Chemaly S., Chung S.W., Liu X, & Perrella M.A. (2021). Mesenchymal stromal cells expressing a dominant‐negative high mobility group A1 transgene exhibit improved function during sepsis. Journal of Leukocyte Biology, 110(4), 711-722.

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Whole Genome Amplification and Aneuploidy Detection

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All procedures were done in accordance with the manufacturer's protocol. PicoPLEX WGA kit (Perkin Elmer) was used to perform whole genome amplification (WGA). The success of amplification was confirmed by assessing the amplified products. Briefly, 2 μL of TE cells' WGA products were amplified with biotin-labelled deoxy-ribonucleoside triphosphate (dNTP) mix for 60-90 min. The samples were hybridised to the KaryoLite BoBs bead set at 52°C in a shaking incubator (800 rpm) for 16 h after removing unbound biotin-labelled dNTPs. Luminex ® 200 TM instrument was used for signal detection. Initial data processing was performed using xPONENT ® software version 3.1.971 (Luminex Corp., Austin, TX, USA). The cvs data file generated was then imported to BoBsoft ® v2.1 (Perkin Elmer) for aneuploidy detection.

Chen N.Q., Si C.R., Yung S.C., Hon S.K., Arasoo J, & Ng S.C. (2024). Analysis of a preimplantation genetic test for aneuploidies in 893 screened blastocysts using KaryoLite BoBs: a single-centre experience. Singapore medical journal.

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Neuroinflammation Assay with BV2 Microglial Cells

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For neuroinflammation assays, BV2 microglial cells were pre-treated with compounds at non-toxic concentrations for 1 h and activated with 500 ng/mL LPS for 24 h. All the experiments were carried out as previously described 19 .
The levels of IL-6, TNF-α and IL-10 were evaluated with a ProCartaPlex Multiplex kit (Thermo Fisher Scientific), following manufacturer's instructions. Luminex 200 TM instrument and xPONENT ® software (LuminexCorp, Austin, TX) were used to collect the data.
The levels of NO were determined with Griess Reagent Kit, which evaluates the spontaneous oxidation of NO to nitrite under physiological conditions. Cells were cultured in DMEM without phenol red at 1x10 6 cells per well in 12-well plates and treated as described before. Then, 150 µL of medium were mixed with 20 µL of Griess reagent and 130 µL of deionized water. Samples were incubated during 30 min at room temperature and the absorbance was measured at 548 nm in a plate reader. All the experiments were performed three times by duplicate.

Alvariño R., Alonso E., Tabudravu J.N., Pérez-Fuentes N., Alfonso A, & Botana L.M. (2021). Tavarua Deoxyriboside A and Jasplakinolide as Potential Neuroprotective Agents: Effects on Cellular Models of Oxidative Stress and Neuroinflammation. ACS chemical neuroscience, 12(1).

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Luminex-based Cytokine Quantification

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Luminex xMAP immunoassay was performed in the University of California, Los Angeles (UCLA) Immune Assessment Core to quantify cytokine levels in the gastrocnemius protein homogenate. Mouse magnetic cytokine/chemokine kits were purchased from EMD Millipore and used per the manufacturer's instructions. Briefly, 25 μl diluted (1:2) samples were mixed with 25 μl magnetic beads and allowed to incubate overnight at 4°C while shaking. After washing the plates twice with wash buffer in a Biotek ELx405 washer, 25 μl of biotinylated detection antibody was added and the samples were incubated for 1 h at room temperature. Then, 25 μl streptavidin-phycoerythrin conjugate was added to the reaction mixture and incubated for another 30 min at room temperature. Following two washes, the beads were resuspended in sheath fluid, and fluorescence was quantified using a Luminex 200TM instrument (Luminex Corporation, TX, USA). Samples presenting cytokine levels below the Luminex detection limit were removed from the analysis.

Oliveira-Santos A., Dagda M., Wittmann J., Smalley R, & Burkin D.J. (2023). Vemurafenib improves muscle histopathology in a mouse model of LAMA2-related congenital muscular dystrophy. Disease Models & Mechanisms, 16(6), dmm049916.

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Multiplexed Glycolysis Pathway Profiling

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The MILLIPLEX^® MAP Human Glycolysis Pathway Magnetic Bead Panel (HGPMAG-27K, Millipore, Burlington, MA, USA) was applied in 96-well plates for the simultaneous quantification of the following proteins in tissue lysates: G6PI (glucose-6-phosphate isomerase), LDHA (L-lactate dehydrogenase A chain), LDHB (L-lactate dehydrogenase B chain), PKM2 (pyruvate kinase isoform M2), TKT (transketolase) and HIF-1α (hypoxia-inducible factor 1-α).
For the immunoassay procedures, 25 µL of each dilute lysate sample in assay buffer (5 µg total protein/well) and HeLa cell lysate (positive control) were added into wells in duplicate, according to the manufacturer’s instructions. To each well, 25 µL of the Mixed Beads were added and the plate was incubated for 2 h at room temperature. Human glycolysis pathway detection biotinylated antibodies were added for 1 h; each captured a specific bead. After that, the reaction mixture was incubated for 30 min with Streptavidin-PE conjugate to complete the reaction on the surface of each microsphere. Finally, the MILLIPLEX^® MAP was analyzed by Luminex xMAP^® technology. The immunoassay on the surface of each fluorescent-coded magnetic bead, MagPlex-C microsphere, was identified and quantified based on fluorescent signals. The median fluorescence intensity (MFI) was read with the Luminex 200TM instrument and measured with xPONENT^® software.

Lucarelli G., Rutigliano M., Loizzo D., di Meo N.A., Lasorsa F., Mastropasqua M., Maiorano E., Bizzoca C., Vincenti L., Battaglia M, & Ditonno P. (2022). MUC1 Tissue Expression and Its Soluble Form CA15-3 Identify a Clear Cell Renal Cell Carcinoma with Distinct Metabolic Profile and Poor Clinical Outcome. International Journal of Molecular Sciences, 23(22), 13968.

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