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Bio plex 100 system

Manufactured by Bio-Rad
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The Bio-Plex 100 system is a multiplex assay platform that enables the simultaneous detection and quantification of multiple analytes in a single sample. The system utilizes fluorescent-encoded beads, flow cytometry, and powerful data analysis software to provide high-throughput, real-time measurements of various biomolecules such as proteins, peptides, and nucleic acids.

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Lab products found in correlation

Bio plex pro human cytokine screening panel 48 plex, by Bio-Rad (3 mentions) Anti human igg biotin, by Merck Group (3 mentions) Magplex magnetic carboxylated microspheres, by DiaSorin (2 mentions) Guava flow cytometer, by Merck Group (2 mentions) Mmyomag 74k, by Merck Group (2 mentions) Multiplex cytokine kit, by Bio-Rad (1 mentions) Bio plex pro rat cytokine 24 plex assay, by Bio-Rad (1 mentions) Sample diluent, by Bio-Rad (1 mentions) Magplex, by DiaSorin (1 mentions) Milliplex human cytokine chemokine panel, by Merck Group (1 mentions) Bio plex manager software 5, by Bio-Rad (1 mentions) Bio plex protm human chemokine panel 40 plex, by Bio-Rad (1 mentions) Minicollect serum separator tubes, by Greiner (1 mentions) Ultra sensitive mouse insulin elisa kit, by Crystal Chem (1 mentions) Bio plex manager 6, by Bio-Rad (1 mentions) Procartaplex, by Thermo Fisher Scientific (1 mentions) Milliplex map human cytokine chemokine magnetic bead panel, by Merck Group (1 mentions) Prism software, by GraphPad (1 mentions)

18 protocols using bio plex 100 system

1

Quantifying IL-1β in Cell Cultures

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Protein levels of IL-1β were measured in culture supernatants using Luminex multianalyte technology on the Bio-Plex 100 system and multiplex cytokine kit (Bio-Rad, Hercules, CA, USA), the sensitivity of which was < 1 pg/mL.
Ascone G., Cao Y., Jansen I.D., Di Ceglie I., van den Bosch M.H., Blom A.B., van Lent P.L., Everts V, & de Vries T.J. (2020). Increase in the Number of Bone Marrow Osteoclast Precursors at Different Skeletal Sites, Particularly in Long Bone and Jaw Marrow in Mice Lacking IL-1RA. International Journal of Molecular Sciences, 21(11), 3774.
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2

Multiplex Cytokine and Chemokine Profiling

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Circulating cytokine and chemokine levels were determined in serum using commercially available bead-based multiplex assays using the BioPLex 100 system for acquisition as previously described.20 Standard curves were included in the kits and, in addition, a pooled serum sample of 4 hospital admitted COVID-19 patients was included as internal reference in all assays. Four commercially available kits were used: Bio-Plex Pro™ Human Cytokine Screening Panel 48-plex, Bio-Plex Protm Human Chemokine Panel 40-Plex, Bio-Plex Protm Human Inflammation Panel 1 and 37-Plex; Bio-Plex Protm Human Th17 panel (all obtained from Bio-Rad, Veenendaal, The Netherlands).
Pongracz T., Nouta J., Wang W., van Meijgaarden K.E., Linty F., Vidarsson G., Joosten S.A., Ottenhoff T.H., Hokke C.H., de Vries J.J., Arbous S.M., Roukens A.H, & Wuhrer M. (2022). Immunoglobulin G1 Fc glycosylation as an early hallmark of severe COVID-19. EBioMedicine, 78, 103957.
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3

Multiplex Antibody Profiling using Luminex

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Measurement of plasma IgG antibodies was performed by multiplex bead-based assay using the Luminex technology, as described (Rui et al., 2011 (link)). The different recombinant proteins and peptides included in this study were covalently coupled to four MagPlex magnetic carboxylated microspheres (Luminex Corporation, TX, USA). Next, in 1.25 million coated beads we coupled 1 μg of each antigen following manufacturer’s instructions. Using a Neubauer chamber coated-beads were quantified and mixed in equal amounts. Only one batch of microspheres was prepared for the study. Plasma (1:100 dilution) was incubated with around 2000 beads per analyte in duplicates, followed by anti-human IgG-biotin (1:4000 dilution) (Sigma-Aldrich) incubation. Next, streptavidin-conjugated to R-phycoerthrin (R-PE) (1 μg/mL) was incubated for 10 min at RT and beads were acquired on the BioPlex100 system (Bio-Rad, Hercules, CA) and express the results as median fluorescence intensity (MFI) of duplicates. Cross-reactivity was ruled out analyzing a subset of plasmas in singleplex and multiplex. A panel of eight negative controls from healthy individuals from Barcelona was included on every plate. Cutoffs for antibody positivity were determined for each plate by calculating mean +2 standard deviation of the negative control values.
Aparici-Herraiz I., Gualdrón-López M., Castro-Cavadía C.J., Carmona-Fonseca J., Yasnot M.F., Fernandez-Becerra C, & del Portillo H.A. (2022). Antigen Discovery in Circulating Extracellular Vesicles From Plasmodium vivax Patients. Frontiers in Cellular and Infection Microbiology, 11, 811390.
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4

Multiplex IgG Detection in MIMIC Supernatants

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Culture supernatants from MIMIC® studies were collected on day 13 for detection of anti-pre-F, anti-Gacc, and anti-Gbcc IgG by a multiplexed Luminex-based method known as Antibody Forensics. Fluorescent carboxylated magnetic beads (Luminex) were activated and coated directly with 2 µg/mL DS-Cav-1 protein, or the peptide corresponding to the RSV Gcc from subgroup A2 or B1 in PBS. The antigen-loaded Luminex beads were thereafter incubated with MIMIC® supernatants and analyzed on the Bioplex-100 system (Bio-Rad).
Rainho-Tomko J.N., Pavot V., Kishko M., Swanson K., Edwards D., Yoon H., Lanza L., Alamares-Sapuay J., Osei-Bonsu R., Mundle S.T., Murison D.A., Gallichan S., Delagrave S., Wei C.J., Zhang L, & Nabel G.J. (2022). Immunogenicity and protective efficacy of RSV G central conserved domain vaccine with a prefusion nanoparticle. NPJ Vaccines, 7, 74.
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5

Multiplex Cytokine Profiling of Rat Serum

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Serum cytokine levels were determined using the Bio-Plex Pro™ Rat Cytokine 24-Plex Assay (Catalog No. 171K1001M; Bio-Rad, Hercules, CA, USA) following the manufacturer’s protocol. In brief, after thawing, serum was centrifuged again at 10 000 × g for 10 min at 4 °C to remove precipitates and platelets. Samples were diluted 1:4 in Bio-Rad sample diluent. Plate was read at High RP1 (PMT) setting using the Bio-Plex 100 system (Bio-Rad, Hercules, CA, USA). A complete list of cytokines tested is found in Table 2.
Russell A.L., Grimes J.M., Cruthirds D.F., Westerfield J., Wooten L., Keil M., Weiser M.J., Landauer M.R., Handa R.J., Wu T.J, & Larco D.O. (2017). Dietary Isoflavone-Dependent and Estradiol Replacement Effects on Body Weight in the Ovariectomized (OVX) Rat. Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et metabolisme, 49(6), 457-465.
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6

Multiplex Plasma IgG Antibody Measurement

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Measurement of plasma IgG antibodies was performed by multiplex suspension array using the Luminex™ technology, as described before (24 (link)). Briefly, 1.1–1.4 million MagPlex® magnetic-carboxylated microspheres (Luminex Corporation, TX, USA) with different spectral signatures were covalently coated with 3 µg of each protein/peptide, following the manufacturer’s instructions. Protein-coupled beads were quantified in a Guava® Flow Cytometer (Millipore) and mixed in equal amounts. A unique batch of microspheres was prepared for the whole study, including the samples analyzed in IN. Approximately 1,000 beads per analyte were incubated with each plasma (1:100 dilution) in duplicates, and subsequently with antihuman IgG-biotin (Sigma-Aldrich), followed by streptavidin-conjugated R-PE (Fluka, Madrid, Spain). Beads were acquired on the BioPlex100 system (Bio-Rad, Hercules, CA, USA), and results were expressed as median fluorescence intensity (MFI) of duplicates. Value against GST alone was subtracted from correspondent proteins. Raw GST data have been previously published (24 (link)). Cross-reactivity was ruled out in a pilot study analyzing a subset of plasmas in singleplex and multiplex (not shown). Samples in IN were analyzed with identical protocols and instruments.
Requena P., Arévalo-Herrera M., Menegon M., Martínez-Espinosa F.E., Padilla N., Bôtto-Menezes C., Malheiro A., Hans D., Castellanos M.E., Robinson L., Samol P., Kochar S., Kochar S.K., Kochar D.K., Desai M., Sanz S., Quintó L., Mayor A., Rogerson S., Mueller I., Severini C., del Portillo H.A., Bardají A., Chitnis C.C., Menéndez C, & Dobaño C. (2017). Naturally Acquired Binding-Inhibitory Antibodies to Plasmodium vivax Duffy Binding Protein in Pregnant Women Are Associated with Higher Birth Weight in a Multicenter Study. Frontiers in Immunology, 8, 163.
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7

Inflammatory Biomarkers in Microdialysis and Plasma

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Microdialysis fluid from the AAR and control Cx region and EDTA-plasma was assessed for inflammatory mediators interleukin (IL)-1β, IL-6, IL-8, IL-10, and TNF using a porcine multiplex cytokine assay on a Bio-Plex 100 system (Bio-Rad, Hercules, CA, USA) as previously described [9 (link)]. LTB4 from plasma and myocardial tissue was measured using a competitive enzyme immunoassay according to the manufacturer’s instructions (R&D systems, Minnesota, MN, USA).
Pischke S.E., Gustavsen A., Orrem H.L., Egge K.H., Courivaud F., Fontenelle H., Despont A., Bongoni A.K., Rieben R., Tønnessen T.I., Nunn M.A., Scott H., Skulstad H., Barratt-Due A, & Mollnes T.E. (2017). Complement factor 5 blockade reduces porcine myocardial infarction size and improves immediate cardiac function. Basic Research in Cardiology, 112(3), 20.
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8

Multiplex Cytokine and Chemokine Profiling

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Concentrations of cytokines and chemokines were determined using the 39-plex kit of MILLIPLEX Human Cytokine/Chemokine Panel (Millipore) and samples were analyzed undiluted on a Bioplex 100 system with Bioplex Manager Software 5.0 (Biorad, Hercules, CA). The cytokines/chemokines detected in this kit includes: interleukin (IL)-1α, IL-1β, IL-1receptor antagonist (RA), IL-2, IL-3, IL-4, IL-5, IL-6, IL-7-IL-8, IL-9, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17, interferon (IFN)γ, IFNα 2, granulocyte-macrophage (GM) colony stimulating factor (CSF), granulocyte (G)-CSF, tumor necrosing factor (TNF) α, TNF β, transforming growth factor (TGF) α, soluble (s)CD40 ligand (L), vascular endothelial growth factor (VEGF), C-C motif chemokine ligand (CCL)-3/ macrophage inflammatory protein (MIP)-1 α, CCL-4/ MIP-1 β, CCL-22/macrophage-derived chemokine (MDC), CCL-2/monocyte chemotactic protein (MCP)-1, CCL7/MCP-3, CXC Motif Chemokine Ligand (CXCL)-10/ interferon gamma-induced protein (IP)-10, CXCL-1/ growth regulated oncogene (GRO), CX3CL-1/ fractalkine, CCL-11/ eotaxin, Fms-like tyrosine kinase(Flt)-3 ligand, epidermal growth factor (EGF), EGF-2, and sCD 25 (sIL-2 receptor).
Hel Z., Huijbregts R.P., Xu J., Nechvatalova J., Vlkova M, & Litzman J. (2014). Altered serum cytokine signature in common variable immunodeficiency. Journal of clinical immunology, 34(8), 971-978.
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9

Multiplex Analysis of Serum Chemokines, Cytokines, and Myokines

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Serum from single-housed 11-month-old males was thawed and applied to multiplex assays for detection of serum chemokines, cytokines (Thermo Fisher Scientific, Cat# EPX360-26092-901, RRID: AB_2576123) and myokines (Milliplex, MMYOMAG-74K, Merck, Kenilworth, NJ, USA). Multiplex microplates were analyzed using a Bio-Plex 100 system (Bio-Rad) according to manufacturer’s instructions. See full list of analytes in Online Resource Supplementary Table S1. Concentrations were determined based on the average of technical duplicates and wells with bead count of less than 20 were excluded to ensure high precision. In cases where the concentrations were below the detection range, concentration value was set as midpoint between 0 and minimum detection limit of the analysis. In the multiplex protein analysis data of the study, sedentary animals from a previous study have been included as data points in the graphs for baseline purpose [35 (link)].
Karlsson L., González-Alvarado M.N., Motalleb R., Wang Y., Wang Y., Börjesson M., Zhu C, & Kuhn H.G. (2020). Constitutive PGC-1α Overexpression in Skeletal Muscle Does Not Contribute to Exercise-Induced Neurogenesis. Molecular Neurobiology, 58(4), 1465-1481.
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10

Cytokine and Chemokine Profiling in COVID-19

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In total 333 sera from 102 COVID-19 positive patients were available for cytokine/chemokine measurements. Cytokines and chemokines were measured in serum by bead based multiplex assays using the BioPLex 100 system (Bio-Rad, Hercules, CA, USA) for acquisition as previously described [28 (link)]. Standard curves were provided with kits and a pooled sample of 4 COVID-19 patients was included as internal reference in all assays. Four commercially available kits were used Bio-Plex Pro™ Human Cytokine Screening Panel 48-plex; Bio-Plex Protm Human Chemokine Panel 40-Plex; Bio-Plex Protm Human Inflammation Panel 1, 37-Plex; Bio-Plex Protm Human Th17 panel (IL-17F, IL-21, IL-23, IL-25, IL-31, IL-33) (all from Bio-Rad, Veenendaal, The Netherlands).
Zlei M., Sidorov I.A., Joosten S.A., Heemskerk M.H., Myeni S.K., Pothast C.R., de Brouwer C.S., Boomaars-van der Zanden A.L., van Meijgaarden K.E., Morales S.T., Wessels E., Janse J.J., Goeman J.J., Cobbaert C.M., Kroes A.C., Cannegieter S.C., Roestenberg M., Visser L.G., Kikkert M., Feltkamp M.C., Arbous S.M., Staal F.J., Ottenhoff T.H., van Dongen J.J., Roukens A.H, & de Vries J.J. (2022). Immune Determinants of Viral Clearance in Hospitalised COVID-19 Patients: Reduced Circulating Naïve CD4+ T Cell Counts Correspond with Delayed Viral Clearance. Cells, 11(17), 2743.
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