Cytokine levels in lung tissues were analyzed using the Bio-Plex protein array system (Bio-Rad Laboratories, Hercules, CA). Briefly, lung tissue was excised and homogenized in 2 ml of ice-cold PBS containing 1× Pierce protease inhibitor cocktail (Thermo Fisher Scientific, Rockford, IL). After homogenization of the lung tissue, Triton X-100 was added to a final concentration of 0.05%, and the samples were clarified by centrifugation. Supernatant fractions from the pulmonary homogenates were then assayed using the Bio-Plex Pro Mouse Cytokine 23-Plex (Bio-Rad Laboratories) for the presence of interleukin 1α (IL-1α), IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-17A, granulocyte colony-stimulating factor (G-CSF), granulocyte monocyte colony-stimulating factor (GM-CSF), gamma interferon (IFN-γ), chemokine (C-X-C motif) ligand 1 (CXCL1)/keratinocyte-derived chemokine (KC), chemokine (C-C motif) ligand 2 (CCL2)/monocyte chemotactic protein 1 (MCP-1), CCL3/macrophage inflammatory protein 1α (MIP-1α), CCL4/MIP-1β, CCL5/regulated upon activation, normal T cell expressed and secreted (RANTES), and tumor necrosis factor alpha (TNF-α).
Bio plex pro mouse cytokine 23 plex
Manufactured by Bio-Rad
Sourced in United States
The Bio-Plex Pro Mouse Cytokine 23-plex is a multiplex assay designed to simultaneously quantify 23 different mouse cytokines and chemokines. It utilizes magnetic bead-based technology to enable the rapid and accurate measurement of multiple analytes from a small sample volume.
Automatically generated - may contain errors
Lab products found in correlation
Pierce protease inhibitor cocktail, by Thermo Fisher Scientific (3 mentions)
Bio plex protein array system, by Bio-Rad (3 mentions)
Bio plex luminex 200, by Bio-Rad (3 mentions)
Bio plex 200, by Bio-Rad (3 mentions)
Ifn betamouse procartaplex simplex kit, by Thermo Fisher Scientific (2 mentions)
Il 28b ifn lambda 3 mouse elisa kit, by Thermo Fisher Scientific (2 mentions)
Bio plex pro tgf β assay, by Bio-Rad (2 mentions)
T per, by Thermo Fisher Scientific (2 mentions)
Protease inhibitor cocktail, by Roche (2 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Duoset elisa, by R&D Systems (1 mentions)
Protease inhibitor, by Thermo Fisher Scientific (1 mentions)
Prism v5, by GraphPad (1 mentions)
Na3vo4, by Cell Signaling Technology (1 mentions)
Np 40, by Cell Signaling Technology (1 mentions)
Enzyme linked immunosorbent assay kit, by Thermo Fisher Scientific (1 mentions)
Millipore filter, by Merck Group (1 mentions)
Bio plex pro mouse chemokine panel 31 plex, by Bio-Rad (1 mentions)
Bio plex 200 system, by Bio-Rad (1 mentions)
Complete protease inhibitor cocktail, by Roche (1 mentions)
25 protocols using bio plex pro mouse cytokine 23 plex
1
Quantifying Cytokine Profiles in Lung Tissue
2
Cytokine Profiling in Lung Tissue
3
Cytokine Profiling in Mice Tissues
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Mice hippocampal tissue (10 mg) or plasma (100 µl) were collected for ELISA. Levels of cytokines in plasma and hippocampal tissues were measured using Bioplex suspension chip reagent Bio-Plex Pro Mouse Cytokine 23-plex (M60009RDPD, Biorad, USA) and normalized with protein concentration in samples.
4
Quantifying Mouse Cytokine Profiles
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Concentrations of mouse cytokines/chemokines were measured in culture media by Bio-Plex Pro™ Mouse Cytokine 23-plex, according to manufacturer’s instructions (Bio-Rad Laboratories Ltd, CA, USA). CCL3 was excluded from the analysis as the samples fell outside of the standard curve.
5
Quantification of TGFβ and IL-6 in Tumor Cells
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TGFβ and IL-6 were measured in the supernatants of tumor cells. In brief, digested tumors were seeded in 96-flat bottom plate. After overnight, the supernatants were collected and stored at −80°C until measurement. Total TGFβ levels were determined using commercially available DuoSet ELISAs (R&D systems, USA) after acidification of the samples to activate latent TGFβ as described before.21 22 (link) IL-6 was measured using a mouse Bio-Plex Pro Mouse Cytokine 23-plex immunoassay (Bio-Rad, Herculus, California, USA) according to manufacturer’s protocol.
6
Multiplex Cytokine Analysis in Murine Sera
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The Bio-Rad Bio-Plex Pro Mouse Cytokine 23-plex was used to test cytokine levels in 12.5 μl of serum according to manufacturer’s instructions. Sera collected from mice 1–4 dpi were analyzed and compared to controls using Kruskal Wallis non-parametric tests. Statistical analyses are described in the figure legend. Cytokine abbreviations: interleukin (IL), keratinocyte chemoattractant (KC), granulocyte-colony stimulating factor (G-CSF), granulocyte/macrophage-colony stimulating factor (GM-CSF), interferon gamma (IFNγ), tumor necrosis factor alpha (TNFα), chemokine ligand (CCL), MCP-1: monocyte chemoattractant protein-1 (CCL2), MIP-1α/β: macrophage inflammatory protein (CCL3/4), RANTES: regulated on activation, normal T cell expressed and secreted (CCL5).
7
Serum Cytokine Profiling of Murine Viral Infection
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Individual serum samples were collected and Bio-Rad Bio-Plex Pro Mouse Cytokine 23-plex used to quantify cytokine levels in 15 ul of serum according to manufacturer’s instructions. Sera collected from mice on days 1–3 pi were analyzed and compared to same-day-matched samples collected from animals inoculated with an equivalent volume of virus-free concentrated tissue culture medium.
8
Cytokine and Chemokine Profiling of EVLP Perfusate
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The perfusates collected after 4 h of EVLP were flash frozen in liquid nitrogen and stored at −80 °C. We assayed 50 µL of perfusate for cytokines, chemokines and mediators of wound healing and tissue repair levels with a mouse cytokine/chemokine panel (Bio-Plex Pro Mouse Cytokine 23-plex, Bio-Rad Laboratories, Hercules, CA, USA) according to the manufacturer’s instructions.
9
Cytokine Production in Pulmonary Phagocytes
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Each subset of pulmonary macrophages or DCs was separated (as described above), and 2x105 cells/ml of each phagocyte subset were incubated with 1µg/ml anti-GXM antibody and 1x104 cells/ml C. neoformans strain H99 (20:1 ratio) in 100 µl in triplicate wells for 2 hours at 37°C, 5% CO2. This time point was chosen to examine early cytokine production following phagocyte-cryptococcal interaction. Following centrifugation, cell supernatants were collected, and protease inhibitor (Thermo Scientific) was added to supernatants. Treated supernatants were placed in tubes and stored at -80°C until analysis. Supernatants were assayed for mouse cytokines/chemokines according to manufacturer’s instructions using the Bio-Plex Pro Mouse Cytokine 23-plex (BioRad, Hercules, CA) and samples were read on a BioRad BioPlex®-200 (Bio-Rad). Data were analyzed using GraphPad Prism v5.0 (GraphPad Software, San Diego, CA).
10
Cytokine Profiling of Renal Cortex
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Renal cortex samples were lysed in lysis buffer containing 150‐mM NaCl, 1‐mM EGTA, 1‐mM EDTA, 20‐mM Tris–HCl, 1‐mM NaF, 1‐mM DTT, 10 μg·μl−1 of aprotinin, 1 μg·μl−1 of leupeptin, 1‐mM Na3VO4, 1‐mM phenylethylmalonylurea fluoride, and 1% NP‐40 (Cell Signaling). Samples were then centrifuged at 20,000 x g for 15 min at 4°C, and the supernatant was collected and stored at −80°C.
Levels of cytokine in the lysates from renal cortex samples were measured by Luminex Assay (# M60009RDPD, Bio‐Plex Pro Mouse Cytokine 23‐plex, Bio‐Rad) according to the manufacturer's protocol.
Levels of cytokine in the lysates from renal cortex samples were measured by Luminex Assay (# M60009RDPD, Bio‐Plex Pro Mouse Cytokine 23‐plex, Bio‐Rad) according to the manufacturer's protocol.
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