Bio plex manager

Manufactured by Bio-Rad

Sourced in United States, Japan

Bio-Plex Manager is a software platform designed to manage and analyze data generated by Bio-Rad's Bio-Plex multiplex assay system. The software provides tools for data acquisition, analysis, and reporting.

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Lab products found in correlation

55 protocols using bio plex manager

Multiplex Cytokine Profiling in Mice and Humans

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Cytokine measurements were performed in duplicates using either the Bioplex 23-plex mouse cytokine kit or the Bioplex 27-plex human cytokine kit from BioRad as per manufacturer's instructions. For mouse BAL fluid analysis, undiluted BAL was mixed with 20% BSA to a final concentration of 1% before use in the bioplex assay.
For human serum samples, 12.5 uL of serum was used for each measurement. The standard curves were optimized automatically by the software (Bioplex manager) and verified manually. The Bioplex manager software was used to calculate cytokine concentrations and only measurements that showed a coefficient of variability (CV) of <10% were included for further analysis. Levels of most cytokines were below detection limit in BAL obtained from Day-0 infected mice as well as in some of the late time points. For the purpose of analysis the values here were set to the lower limit of detection in each case.

Kumar Y., Liang C., Limmon G.V., Liang L., Engelward B.P., Ooi E.E., Chen J, & Tannenbaum S.R. (2014). Molecular Analysis of Serum and Bronchoalveolar Lavage in a Mouse Model of Influenza Reveals Markers of Disease Severity That Can Be Clinically Useful in Humans. PLoS ONE, 9(2), e86912.

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Detecting Bronchiolitis Obliterans Progression

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There were two primary end-points: 1) a composite of BOS progression-free survival, defined as time from randomisation to ≥20% decline in FEV₁, re-transplantation or death, whichever occurred first (prolonged mechanical ventilation and irreversible respiratory failure equivalent to ≥20% decline of FEV₁), and 2) BOS grade progression by grade changes from randomisation to study completion. A decline in FEV₁ was validated for absence of concurrent illness measured at intervals ≥3 weeks apart.
Other exploratory end-points included change in lung function, quantitation of histological bronchiolitis obliterans and bronchoalveolar lavage (BAL) cytokine measurements (interleukin (IL)-1β, IL-2, IL-6, IL-8, IL-10, IL-17, interferon (IFN)-γ and tumour necrosis factor (TNF)-α)) by multiplex assay (Luminex 100 system; Luminex, Austin, TX, USA) analysed using Bio-Plex Manager (Bio-Rad, Hercules, CA, USA). BAL was performed before randomisation, at week 24, and when indicated clinically [2 (link)].

Iacono A., Wijesinha M., Rajagopal K., Murdock N., Timofte I., Griffith B, & Terrin M. (2019). A randomised single-centre trial of inhaled liposomal cyclosporine for bronchiolitis obliterans syndrome post-lung transplantation. ERJ Open Research, 5(4), 00167-2019.

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Multiplex Cytokine Profiling in Serum

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A multiplex assay for quantitative determination of inflammatory mediators was applied to assess the concentrations of selected cytokines in serum. The analysis was performed using the Bio-Plex Pro Human Cytokine 17-Plex panel (Bio-Rad Laboratories GmbH). The following determinants were simultaneously detected: IL-1β, IL-2, IL-4, IL-5, IL6, IL-7, IL-8, IL-10, IL-12, IL-13, IL-17, G-CSF, GM-CSF, IFN-γ, MCP-1, MIP-1β and TNF-α. Multiplexing was performed according to the manufacturer’s instruction manual and analyzed on a Bio-Rad Bio-Plex 200 system. Values were calculated by the Bio-Plex^™ software (Bio-Plex Manager, version 6.1, Bio-Rad). All serum samples were measured in triplicates.

Schimmack S., Yang Y., Felix K., Herbst M., Li Y., Schenk M., Bergmann F., Hackert T, & Strobel O. (2019). C-reactive protein (CRP) promotes malignant properties in pancreatic neuroendocrine neoplasms. Endocrine Connections, 8(7), 1007-1019.

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Measurement of Soluble Immune Checkpoints

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The levels of solCRs in plasma and PCTS supernatants were analysed by multiplex Luminex technology using a commercially available custom 14-plex Immuno-Oncology Checkpoint Human ProcartaPlex Panel (ThermoFisher Scientific) according to the manufacturer’s instructions. The following 14 checkpoints were included in the panel: sBTLA; sCD28; sLAG3; sCD40; sCD80; sCD137; sCD152 (CTLA-4); sGITR; sHVEM; IDO-1; sPD-1; sPD-L1; sPD-L2; sTIM3. Measurements were performed using a Luminex MAGPIX Instrument and analysed using XPonent MAGPIX 4.2 and BioPlex Manager (Bio-rad, version 6). In samples where the solCR concentration was recorded as undetectable, a value corresponding to half the lowest detectable value was used for the purpose of analysis.

Doornebal E.J., Harris N., Riva A., Jagatia R., Pizanias M., Prachalias A., Menon K., Preziosi M., Zamalloa A., Miquel R., Zen Y., Orford M.R., Eaton S., Heaton N., Ramage J., Palma E., Srirajaskanthan R, & Chokshi S. (2022). Human Immunocompetent Model of Neuroendocrine Liver Metastases Recapitulates Patient-Specific Tumour Microenvironment. Frontiers in Endocrinology, 13, 909180.

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Multiplex Cytokine Profiling of Activated pDCs

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Initial screening of cytokine concentrations in the supernatant of the stimulated pDCs was performed using Bio-Plex Pro Human Cytokine 17-Plex Panel (Bio-Rad), complemented by a Bio-Plex Pro Human Cytokine IP-10 (Bio-Rad) and Bio-Plex Pro Human Cytokine IFN-a2 (Bio-Rad) resulting in a total of 19 cytokine targets (IP-10, MIP-1β, TNF-α, IFN-α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL-17, G-CSF, GM-CSF, IFN-γ and MCP-1). Consecutive analyses were performed using a combination of Bio-Plex Pro Human Cytokine IP-10, Bio-Plex Pro Human Cytokine MIP-1α, Bio-Plex Pro Human Cytokine TNF-α and Bio-Plex Pro Human Cytokine IFN-a2 sets (Bio-Rad).
Multiplex assay was performed on a Luminex 200 system (Bio-Rad) in accordance with the manufacturer’s instructions. Standards were analysed in duplicates and each sample in triplicates. Washing was performed between each step using a HydroFlex microplate equipped with a magnetic plate carrier (Tecan). Analysis of the data was performed using Bio-Plex Manager (Bio-Rad).

Bucher K., Rodríguez-Bocanegra E., Wissinger B., Strasser T., Clark S.J., Birkenfeld A.L., Siegel-Axel D, & Fischer M.D. (2023). Extra-viral DNA in adeno-associated viral vector preparations induces TLR9-dependent innate immune responses in human plasmacytoid dendritic cells. Scientific Reports, 13, 1890.

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Pediatric Vaccine Multiplex Assay

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An adaptation of the recently published pediatric vaccine multiplex assay (PVMA) was used to measure maternal and infant antigen-specific IgG levels [29 (link)]. Briefly, 25 μg of antigens were coupled to a total of 5 × 10⁶ carboxylated microspheres (Bio-Rad Laboratories, Hercules, CA), and 5,000 antigen-coupled microspheres of each of the antigens were then added to each well. Standard curves and plasma samples were prepared in duplicate in a background-reducing assay diluent (PBS, 1% evaporated milk, 5% goat serum, 0.05% polyvinyl alcohol, and 0.08% polyvinylpyrrolidone) and incubated on a microplate shaker at room temperature for 1 hour before added to the wells (Table S1). Antigen-specific IgG was detected with mouse anti-human IgG-PE at 2 μg/mL (SouthernBiotech, Birmingham, AL). The plates were read on a Bio-Plex 200 system (Bio-Rad Laboratories, Hercules, CA). All mean fluorescent intensity (MFI) values were blank well subtracted. Bio-Plex Manager (Bio-Rad Laboratories, Hercules, CA) was used to interpolate unknown concentrations from standard curves using the background-subtracted MFIs.
Intra-assay coefficients of variance (CVs), which are the average CVs of duplicate wells within one assay, were determined for each sample and samples with CVs greater than 25% were repeated. Samples with blank bead MFIs greater than 1,000 were excluded from analysis.

Post A.L., Li S.H., Berry M., Itell H., Martinez D.R., Xie G., Permar S.R., Swamy G.K, & Fouda G.G. (2020). Efficiency of Placental Transfer of Vaccine-elicited Antibodies Relative to Prenatal Tdap Vaccination Status. Vaccine, 38(31), 4869-4876.

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Maternal Serum Cytokine and Endotoxin Quantification

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Maternal serum cytokine concentrations were measured using the Bio-Plex Pro™ Mouse Cytokine 23-plex Assay kit (Bio-Rad Laboratories) following the manufacturer’s instructions. Reading was performed using a Bio-Plex® 200 system (Bio-Rad Laboratories) using Bio-Plex Manager™ software (Bio-Plex Manager™ 6.1 © Copyright 2000 Bio-Rad Laboratories, Inc.).
Maternal serum endotoxin concentrations were measured using the Pierce™ LAL Chromogenic Endotoxin quantitation kit (Thermo Fisher Scientific) following the manufacturer’s instructions. Reading was performed using a plate reader at a wavelength of 405 nm (BioTek®, Oakville, Canada).

Wallace J.G., Bellissimo C.J., Yeo E., Fei Xia Y., Petrik J.J., Surette M.G., Bowdish D.M, & Sloboda D.M. (2019). Obesity during pregnancy results in maternal intestinal inflammation, placental hypoxia, and alters fetal glucose metabolism at mid-gestation. Scientific Reports, 9, 17621.

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Adipokine Profiling in Differentiated ADSCs

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After differentiation ADSCs in adipocytes, the content of the following 10 adipokines, cytokines and chemokines were determined in the incubation medium by the method of flow fluorimetry (multiplex analysis, Luminex xMAP): LEPTIN, ADIPONECTIN, ADIPSIN, VASPIN, CHEMERIN, TNF-α, IL-6, IL-8, IL-10, МСР-1, IP-10. This method included a multiplex immune reaction that took place on the various-diameter microparticles carrying the absorbed antibodies, and subsequent flow fluorescence analysis as well as simultaneous measurement of cytokine concentration. The procedures were conducted according to the Bio-Plex Pro Assay protocol. The results were recorded using an automatic photometer for Bio-Plex microplates (Bio-Plex 200 Systems, Bio-Rad, USA) and the Bio-Plex Manager (“Bio-Rad”) software. The test substances concentration was determined from the calibration curve for each cytokine (the dynamic range 2–32 000 pg/mL) according to the manufacturer recommendations. Detailed description of the methods was presented in previous studies^{46 (link)}.

Zlatska A.V., Vasyliev R.G., Gordiienko I.M., Rodnichenko A.E., Morozova M.A., Vulf M.A., Zubov D.O., Novikova S.N., Litvinova L.S., Grebennikova T.V., Zlatskiy I.A, & Syroeshkin A.V. (2020). Effect of the deuterium on efficiency and type of adipogenic differentiation of human adipose-derived stem cells in vitro. Scientific Reports, 10, 5217.

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Fluorometric Insulin and Adiponectin Assay

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Quantitative determination of insulin and adiponectin in plasma was performed by flow-through fluorometry using commercial test systems (Bio-Rad, USA) on a two-beam laser automated analyzer (BioPlex® 200 Systems, Bio-Rad, USA) and BioPlex Manager (Bio-Rad, USA).

Skuratovskaia D., Zatolokin P., Vulf M., Mazunin I, & Litvinova L. (2019). Interrelation of chemerin and TNF-α with mtDNA copy number in adipose tissues and blood cells in obese patients with and without type 2 diabetes. BMC Medical Genomics, 12(Suppl 2), 40.

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Multiplex Binding Assay for gp140 Antibodies

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A customized multivariate multiplex assay was developed using a panel of gp140 antigens (Clade C: CN54, 1086, Clade A: UG37, Clade D: UG21–NIH AIDS Reagents) covalently conjugated to different magnetic fluorescent multiplex beads (Bio-Rad, AU) as described previously (25 (link)). Biotinylated dimeric Fc-gamma-Receptors (FcγRIIa-H131, FcγRIIIa-V158) were produced as previously described (26 (link)). The dimeric FcγR multiplex method has been previously published (21 (link)). Briefly, gp140 coupled microspheres (minimum 500 of each individual antigen bead set per well) was added to 1:100 plasma diluted in PBS + beads, incubating overnight at 4°C. HIVIG was used as a positive control to normalize across multiple replicates. Beads were washed using a Bio-Rad magnetic plate-washer (Bio-Plex Pro Wash station) and incubated for 2 h with biotinylated dimeric FcγR (1.0 μg/ml), then subsequently washed and incubated 1 h with streptavidin PE (1.0 μg/ml), washed again before resuspending in sheath fluid. A Bio-plex MAGPIX and Bio-Plex Manager software (Bio-Rad) was used to detect the median fluorescence intensity (MFI) for each bead set.

Kratochvil S., McKay P.F., Chung A.W., Kent S.J., Gilmour J, & Shattock R.J. (2017). Immunoglobulin G1 Allotype Influences Antibody Subclass Distribution in Response to HIV gp140 Vaccination. Frontiers in Immunology, 8, 1883.

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