Blood samples were collected using EDTA anticoagulation tubes and then centrifuged at 2000 rpm for 10 minutes at room temperature. We used the Bio-Plex 200 system (Bio-Plex Pro Human Cytokine Grp I Panel 27-Plex; Bio-Rad, USA) as specified by the manufacturer of the commercial kit. The procedure is detailed in a previous study.20 (link) In this study, we detected a total of 27 cytokines, including IL-1β, IL-1RA, IL-6, IL-8, IL-17A, IL-10, and TNF-α (immunomodulatory cytokines); FGF-basic, VEGF, G-CSF, and GM-CSF (growth cytokines); IFN-γ, IL-2, IL-4, IL-5, IL-7, IL-13 IL-12p70, IL-15, and IP-10 (helper T cell-associated cytokines); IL-9, MCP-1, RANTES, MIP-1α, MIP-1β, PDGF-BB, and Eotaxin.
Bio plex pro human cytokine grp 1 panel 27 plex
Manufactured by Bio-Rad
Sourced in United States
The Bio-Plex Pro Human Cytokine Grp I Panel 27-Plex is a multiplexed immunoassay that simultaneously quantifies 27 different human cytokines and chemokines in a single sample. The panel includes a comprehensive set of analytes that can be used to profile the human immune response.
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Lab products found in correlation
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Bio plex 200 system, by Bio-Rad (2 mentions)
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Bio plex magpix, by Bio-Rad (1 mentions)
Gm csf, by BD (1 mentions)
Interleukin 6 (il 6), by BD (1 mentions)
Bio plex multiplex immunoassay, by Bio-Rad (1 mentions)
Bio plex magpix system, by Bio-Rad (1 mentions)
Bio plex 200 platform, by Bio-Rad (1 mentions)
14 protocols using bio plex pro human cytokine grp 1 panel 27 plex
1
Multiplex Cytokine Profiling from EDTA Blood
2
Multiplex Cytokine Profiling in Plasma
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Plasma samples were obtained using a commercial kit (Bio-Plex Pro Human Cytokine Grp I Panel 27-Plex; Bio-Rad, USA) on the Bio-Plex 200 System. The detected cytokines included Th1 type cytokines (IL-2, IL-7, IL-12p70, IL-15, IFN-γ, and IP-10), Th2 (IL-4, IL-5, and IL-13), proinflammatory cytokines (IL-1β, IL-1RA, IL-6, IL-8, IL-17A, and TNF-α), an immunoregulatory cytokine (IL-10), growth factors (FGF-basic, VEGF, G-CSF, and GM-CSF), and several other chemokines (IL-9, MIP-1α, MIP-1β, MCP-1, RANTES, Eotaxin, and PDGF-BB). Briefly, a gradient dilution standard was prepared according to the specification, and each standard was thoroughly mixed during the preparation process. The diluted plasma was incubated in plates containing magnetic antibody-coupled beads and incubated under shock for 30 min. Thereafter, the sample was washed and incubated with the secondary antibodies, as described previously herein. Streptavidin-PE was added to the sample at the end of incubation and washing. Bio-plex Manager 5.0 Software was used to obtain the raw data. The concentrations of 27 cytokines were determined using standard curves. Quality control standards were strictly followed during the experiment.
3
Evaluating Cytotoxic Function via CD107 Assay
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The CD107 externalization assay was performed to evaluate cytotoxic function.32 (link) Cells (2×105) were cocultured with the same number of Daudi cells in the presence of 10 μmol/L Monensin (Sigma) and 5 μL of FITC-conjugated anti-CD107a and CD107b antibody (BD) or isotype control (mouse IgG1, BD). After 4 hours incubation, CD107 expression was determined by flow cytometry as an indicator of degranulation of cytotoxic molecules. At the same time, culture supernatants were harvested and analyzed for cytokine and chemokine production using Bio-Plex Pro Human Cytokine Grp I panel 27-plex (BIO-RAD, Hercules, California, USA).
4
Plasma Cytokine Profiling in Leishmaniasis
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5 mL of blood from patients with Lg-CL before antimonial treatment and from healthy controls were collected. Plasma was separated and kept frozen at -80°C until plasma cytokines assay.
The levels of FGF basic, Eotaxin, G-CSF, GM-CSF, IFNγ, IL-1β, IL-1RA, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17A, IP-10, MCP-1, MIP-1α, MIP-1β, PDGF-BB, RANTES, TNFα and VEGF were determined using the multiplex cytokine commercial kit Bio-PlexPro-Human Cytokine GrpI Panel 27-Plex (Bio-Rad) according to the manufacturer’s instructions in the Bio-Plex 200 Protein Array System (Luminex Corporation).
The levels of FGF basic, Eotaxin, G-CSF, GM-CSF, IFNγ, IL-1β, IL-1RA, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17A, IP-10, MCP-1, MIP-1α, MIP-1β, PDGF-BB, RANTES, TNFα and VEGF were determined using the multiplex cytokine commercial kit Bio-PlexPro-Human Cytokine GrpI Panel 27-Plex (Bio-Rad) according to the manufacturer’s instructions in the Bio-Plex 200 Protein Array System (Luminex Corporation).
5
Comparative Cytokine Profiling of CD11c+ and CD11c- B Cells in Graves' Disease
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To determine the difference in cytokine profiles between CD11c+ B cells and CD11c- B cells, five batches of paired CD11c+ B cells and CD11c- B cells from 15 GD patients were seeded in a round-bottom 96-well plate at 2×105 cells/well. Then, the sorted B cells were activated with 10 µg/ml Phaseolus vulgaris agglutinin (PHA; Sigma–Aldrich, USA) and cultured in a 37°C incubator with 5% CO2 for 72 h. Culture supernatant samples for the paired CD11c+ B cells and CD11c- B cells were collected and stored at -80°C for Luminex liquid suspension chip detection, which was conducted by Wayen Biotechnologies (Shanghai, China). The Bio-Plex Pro Human Cytokine Grp I Panel 27-plex (Bio–Rad, USA) was used to quantify the concentrations of cytokines secreted by CD11c+ B cells and CD11c- B cells from GD patients according to the manufacturer’s instructions. The lower limits of detection (LLOD) of all cytokines in the Luminex liquid suspension chip are shown in Supplementary Table S3
6
Plasma Cytokine Profiling in Lg-CL
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5 mL of blood from patients with Lg-CL before antimonial treatment and from healthy controls were collected. Plasma was separated and kept at -80°C until plasma cytokines assay.
The levels of IFNγ, IL-1β, IL-6, IL-8, IL-12 (p70), IL-13, IL-17A, RANTES, and TNFα were determined using the multiplex cytokine commercial kit Bio-PlexPro-Human Cytokine GrpI Panel 27-Plex (Bio-Rad) according to the manufacturer’s instructions in the Bio-Plex 200 Protein Array System (Luminex Corporation).
The levels of IFNγ, IL-1β, IL-6, IL-8, IL-12 (p70), IL-13, IL-17A, RANTES, and TNFα were determined using the multiplex cytokine commercial kit Bio-PlexPro-Human Cytokine GrpI Panel 27-Plex (Bio-Rad) according to the manufacturer’s instructions in the Bio-Plex 200 Protein Array System (Luminex Corporation).
7
Multiplex Analysis of Cytokine Homeostasis
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Thanks to soluble factors, including cytokines, the immune system is able to maintain homeostasis through activating and inhibiting signals while adjusting the response to environmental signals [11] . Modern platforms such as Bio-Plex Magpix by Bio-Rad allow the simultaneous determination of up to 100 proteins, peptides, and nucleic acids in a single sample [17] .
In this study, each day immediately after the exposure to EMF, supernatants from the test and control groups were collected and frozen at -80°C until the analysis was performed. In total, 30 analytes were measured using 2 kits: Bio-Plex Pro TGF-β Panel 3-Plex (Bio-Rad) and Bio-Plex Pro Human Cytokine Grp I Panel 27-Plex (Bio-Rad).
Data analysis was performed on a Bio-Plex 200 system in conjunction with Bio-Plex Manager ver. 6.1.1 using a 5-parameter (5-PL) nonlinear logistic regression curve fit model (Bio-Rad). According to the Bio-Rad Bio-Plex Multiplex Immunoassay Instruction the sensitivity of the assay was defined as analyte concentration corresponding to the median background fluorescence intensity (MFI) plus 2 standard deviations (SD) of the mean background MFI.
In this study, each day immediately after the exposure to EMF, supernatants from the test and control groups were collected and frozen at -80°C until the analysis was performed. In total, 30 analytes were measured using 2 kits: Bio-Plex Pro TGF-β Panel 3-Plex (Bio-Rad) and Bio-Plex Pro Human Cytokine Grp I Panel 27-Plex (Bio-Rad).
Data analysis was performed on a Bio-Plex 200 system in conjunction with Bio-Plex Manager ver. 6.1.1 using a 5-parameter (5-PL) nonlinear logistic regression curve fit model (Bio-Rad). According to the Bio-Rad Bio-Plex Multiplex Immunoassay Instruction the sensitivity of the assay was defined as analyte concentration corresponding to the median background fluorescence intensity (MFI) plus 2 standard deviations (SD) of the mean background MFI.
8
Cytokine and Chemokine Profiling in SARS-CoV-2
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Supernatants from SARS-CoV-2 experiments were collected for measurement of the chemokine IL-8 (BD) and the cytokines IL-6 (BD) and GM-CSF (BD) by ELISA, following the manufacturer’s instructions. High throughput cytokine/chemokine measurement was done by using Bio-Plex Pro™ Human Cytokine Grp I Panel 27-Plex (Bio-Rad), following the manufacturer’s instruction.
9
Multiplex Cytokine and Chemokine Profiling
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Serum cytokines concentration: interleukins-1b, -2, -4, -5, -6, -7, -8 (CXCL8), -9, -10, -12, -13, -15, -17A, TNF-α, IFN-γ, TGF-β1-3) chemokines (MIP-1a (CCL3), MIP-1b (CCL4), MCP-1, RANTES (CCL5), Eotaxin (CCL11), IP-10) growth factors (G-CSF, GM-CSF, PDGF, bFGF and VEGF) and receptor antagonist of IL-1 were measured using Bio-Plex Pro TGF-β Panel 3-Plex (Bio-Rad) and Bio-Plex Pro Human Cytokine Grp I Panel 27-Plex (Bio-Rad, Poland). Data analysis was performed on a Bio-Plex 200 system in conjunction with Bio-Plex Manager version 6.1.1 using a 5-parameter (5-PL) nonlinear logistic regression curve fit model (Bio-Rad). As per Bio-Rad Bio-Plex Multiplex Immunoassay recommendations, the assay's sensitivity was defined as analyte concentration corresponding to the median background fluorescence intensity (MFI) plus two standard deviations (SD) of the mean background MFI.
10
Comprehensive Cytokine Profiling in Plasma
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Plasma was collected in the process of PBMC isolation and frozen in small aliquots at −80°C and subjected to measurement of 27 proteins (IL-1ra; IL-1β; IL-2; IL-4; IL-5; IL-6; IL-7; IL-8; IL-9; IL-10; IL-12p70; IL-13; IL-15; IL-17A; C-C motif chemokine ligand 11 [CCL11; Eotaxin]; fibroblast growth factor 2 [FGF-2]; colony stimulating factor 3 [CSF3; G-CSF]; colony stimulating factor 2 [CSF2; GM-CSF]; interferon gamma [IFN-γ]; tumor necrosis factor alpha [TNF-α]; C-X-C motif chemokine ligand 10 [CXCL10; IP-10]; C-C motif chemokine ligand 2 [CCL2; MCP-1]; C-C motif chemokine ligand 3 [CCL3; MIP-1α]; C-C motif chemokine ligand 4 [CCL4; MIP-1β]; platelet-derived growth factor-BB [PDGF-BB]; regulated on activation, normal T cell expressed and secreted [RANTES]; and vascular endothelial growth factor [VEGF]). These were measured with Bio-Plex Pro Human Cytokine Grp I Panel 27-plex (BIO RAD, Hercules, CA, USA). Arginase and iNOS were also quantified with enzyme-linked immunosorbent assays (Hycult Biotech, Uden, Netherlands and Cloud-Clone Corp, Houston, TX, USA, respectively).
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