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Bio plex 200 system instrument

Manufactured by Bio-Rad
Sourced in United States

The Bio-Plex 200 system is a multiplex assay instrument designed to perform simultaneous quantification of multiple analytes in a single sample. The system utilizes xMAP technology to enable high-throughput analysis of a wide range of biomolecules, including proteins, cytokines, and nucleic acids. The instrument is capable of detecting and analyzing up to 100 different analytes in a single reaction.

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8 protocols using bio plex 200 system instrument

1

TGF-β1 Quantification in Feces

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The concentration of TGF-β1 in the feces was determined by using a Bio-Plex 200 system instrument and the Bio-Plex ProTM TGF-β Assay (both from Bio-Rad, Hercules, CA, USA). Analysis of samples was carried out as previously described [17 (link)].
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2

Multiplex Cytokine Quantification in Monkeys

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Multiplex cytokine quantification was performed using the Cytokine 29-Plex Monkey Panel kit (Invitrogen Catalogue number LPC0005M; ThemoFisher Scientific) according to manufacturer’s instructions and as previously described [25 (link)]. In brief, EDTA-anticoagulated blood samples were centrifuged (14000×g for 5 min) and plasma aliquots were cryopreserved at −80 °C until used. Once-thawed plasma samples were filtered through an Ultrafree Centrifugal Filter (Millipore Sigma) just prior to assay. The reaction plates were read on a Bioplex-200 system instrument and the results were calculated using BioPlex software version 6.2 (BioRad, Hercules, CA).
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3

Multiplex Cytokine and Chemokine Profiling

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Plasma cytokines and chemokines, including IL-1β, IL-1 receptor antagonist, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12p70, IL-13, IL-15, IL-17A, eotaxin, fibroblast growth factor (FGF)-basic, granulocyte colony-stimulating factor, granulocyte macrophage colony-stimulating factor (GM-CSF), IFN-γ, IP-10, monocyte chemoattractant protein-1, macrophage inflammatory protein (MIP)-1α, MIP-1β, platelet-derived growth factor (regulated on activation, normal T cell expressed and secreted), tumor necrosis factor alpha (TNF-α), and vascular endothelial growth factor were measured using the Bio-Plex Pro Human Cytokine Standard 27-Plex Assays panel and the Bio-Plex 200 system instrument (Bio-Rad, Hercules, CA, USA) according to the manufacturer's instructions [19 (link),20 (link)].
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4

Multiplex Plasma Cytokine Profiling

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Twenty-seven plasma cytokines, chemokines, and growth factors were analyzed using the Bio-Plex Pro Human Cytokine 27-plex panel (Bio-Rad Laboratories Inc, Hercules, CA, USA) following the manufacturer instructions. Prior to quantification, plasma samples were diluted 1 : 4 in standard diluent (provided by the manufacturer). By using such panel, serum samples were screened for interleukin (IL)-1β, IL-1RA, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12, IL-13, IL-15, IL-17, eotaxin, basic fibroblast growth factor (FGF), granulocyte-colony stimulating factor (G-CSF), granulocyte-macrophage colony stimulating factor (GM-CSF), interferon (IFN)-γ, IFN-γ induced protein-10 (IP-10), monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein (MIP)-1 α/β (MIP-1α, MIP-1β), platelet-derived growth factor (PDGF), RANTES (regulated on activation normal T-cell expressed and secreted), tumor necrosis factor α (TNF-α), and vascular endothelial growth factor (VEGF) concentrations. Fluorescent signals were recorded using a Bio-Plex 200 System instrument and analyzed using the Bio-Plex Manager Software (Bio-Rad Laboratories Inc, Hercules, CA, USA). The software fitted samples' median fluorescence intensity (MFI) values versus standards' MFI and converted it to concentration (pg/mL) by applying a five-parameter logistic regression (as suggested by the manufacturer).
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5

Multiplex Cytokine Quantification

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TNF-α, IL-17, IL-1β, IL-6, and IFN-γ were quantified in serum using a convenient bioplex kit assay, according to the manufacturer’s instructions. The concentrations of cytokines were measured using Bio-Plex™ 200 System instrument (Bio-Rad Laboratories, Hercules, CA, USA) and Bio-Plex manager 4.1 Software.
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6

Multiplex Cytokine Measurement in Samples

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IL-6 and TNF-α were measured using a commercial enzyme-linked immunosorbent assay (ELISA) kit following the manufacturer’s instructions (R&D Systems, Minneapolis, MN, USA). Results were measured at an optical density of 450 nm (reference wavelength 630 nm), and concentrations in pg/mL were derived from the standard curve. IL-1β, IL-10, Regulated upon Activation, Normal T Cell Expressed and Presumably Secreted (RANTES) and MCP-1 were measured using a multiplex bead array kit according to the manufacturer’s instructions (Bio-Rad) with results analyzed on a Bio-Plex 200 system instrument (Bio-Rad) fitted with the Bio-Plex Manager v6 software (Bio-Rad) and results reported in pg/mL.
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7

Measuring Plasma Cytokines and Biomarkers

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Proinflammatory cytokines in plasma were measured by Luminex 200 sytems (Bio-Rad Inc., Hercules, CA, USA) according to the manufacturer’s instructions. Prior to assays, plasma samples were thawed and centrifuged. Cytokine levels were measured using the ProcartaPlex NHP cytokine/GF37plex (Invitrogen) according to manufacturer’s instructions. The reactions in microtiter plates were read on a Bioplex-200 system instrument and results were calculated using BioPlex software version 6 (BioRad, Hercules, CA). Plasma p27 and anti-SIV gp120 were measured with standard ELISA (p27 ELISA kit, Zeptometrix Corp., Buffalo, NY; native SIV gp120, ABL, Rockville, MD).
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8

Biomarker Profiling of Human Milk

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Concentrations of Igs (IgA, IgG, and IgM), transforming growth factor-beta 2 (TGF-β2) and hormones (adiponectin, ghrelin, and leptin) were determined in duplicate using a Bioplex 200 system instrument (Bio-Rad, Hercules, CA, USA) and the Bio-Plex Pro Human Isotyping Assay, Bio-Plex Pro Human TGF-β Assay and Bio-Plex Pro Human Diabetes Assays kits (Bio-Rad). The concentration of EGF was measured using the RayBio® Human EGF ELISA kit (RayBiotech, Norcross, GA). Every assay was performed according to manufacturer's instructions. Standard curves were performed for each analyte. The analytes were assayed in, at least, three batches for each HTST treatment.
The inter-assay coefficients of variation were below manufacturers' instructions for all the immune markers, and the lower limit of quantification (LLOQ) for every analyte in human milk were: 0.21 μg/L for IgA, 0.78 μg/L for IgM, 2.19 μg/L for IgG, 1.57 ng/L for TGF-β2, 0.03 ng/L for EGF, 23.10 ng/L for adiponectin, 7.40 ng/L for ghrelin, and 11.45 ng/L for leptin.
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