Bio plex multiplex cytokine assay

Manufactured by Bio-Rad

Sourced in United States

The Bio-Plex multiplex cytokine assay is a laboratory instrument used for the simultaneous quantification of multiple cytokines in a single sample. The core function of this product is to detect and measure the concentrations of various cytokines, which are signaling proteins involved in immune system regulation and cell-to-cell communication.

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20 protocols using bio plex multiplex cytokine assay

Cytokine Profiling in CSF

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After lumbar puncture, CSF was centrifuged and immediately stored at −80 °C until analysed using a Bio-Plex multiplex cytokine assay (Bio-Rad Laboratories, Hercules, CA, USA) according to the manufacturer’s instructions. CSF concentrations of IL-1β were calculated according to a standard curve generated for the specific target and expressed as picograms per milliliter (pg/mL).

Stampanoni Bassi M., Buttari F., Nicoletti C.G., Mori F., Gilio L., Simonelli I., De Paolis N., Marfia G.A., Furlan R., Finardi A., Centonze D, & Iezzi E. (2020). Interleukin-1β Alters Hebbian Synaptic Plasticity in Multiple Sclerosis. International Journal of Molecular Sciences, 21(19), 6982.

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Multiplex Cytokine Assay in Rats

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Serum cytokine levels were measured in rats treated with or without Ex-4 for 6 weeks. Levels of IL-1β, MCP-1, IL-6, IL-10 and TNFα were simultaneously measured using the Bio-Plex Multiplex Cytokine Assay (Bio–Rad Laboratories) according to the manufacturer's protocol. Samples below detection limit were assigned a value corresponding to half of the sensitivity of the assay (Assay sensitivity: IL-1β, 2pg/ml; MCP-1, 3pg/ml; IL-6, 10 pg/ml and TNFα, 3 pg/ml).

Larsson M., Lietzau G., Nathanson D., Östenson C.G., Mallard C., Johansson M.E., Nyström T., Patrone C, & Darsalia V. (2016). Diabetes negatively affects cortical and striatal GABAergic neurons: an effect that is partially counteracted by exendin-4. Bioscience Reports, 36(6), e00421.

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Cytokine Profiling in Multiple Sclerosis Patients

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CSF was collected by LP. Immediately after LP, CSF was centrifuged and stored at −80°C until being analyzed using a Bio-Plex multiplex cytokine assay (Bio-Rad Laboratories, Hercules, CA, USA), according to the manufacturer’s instructions. CSF detectability of the following molecules has been examined: interleukin (IL)-1β, IL-1 receptor antagonist (IL-1ra), IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12, IL-13, IL-15, IL17, tumor necrosis factor alpha (TNFa), IFNg, macrophage inflammatory protein (MIP)-1α/CCL3, MIP-1β/CCL4, monocyte chemoattractant protein (MCP)-1/CCL2, interferon gamma-induced protein 10 (IP-10)/CXCL10, eotaxin, granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF), platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), vascular-endothelial growth factor (VEGF), and Regulated on Activation, Normal T Cell Expressed and Secreted (RANTES)/CCL5. CSF delectability of all molecules tested in our CIS and RRMS patients, separated in three groups according to DMTs (IFN-beta, GA, DF) used, is presented in the Supplemental Table.

Stampanoni Bassi M., Drulovic J., Pekmezovic T., Iezzi E., Sica F., Gilio L., Gentile A., Musella A., Mandolesi G., Furlan R., Finardi A., Marfia G.A., Bellantonio P., Fantozzi R., Centonze D, & Buttari F. (2020). Cerebrospinal fluid inflammatory biomarkers predicting interferon-beta response in MS patients. Therapeutic Advances in Neurological Disorders, 13, 1756286420970833.

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Cerebrospinal Fluid IL-10 Profiling

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In 73 patients CSF concentrations of IL-10 were analyzed. CSF was collected at the time of diagnosis, during hospitalization, by lumbar puncture (LP). CSF samples were stored at −80°C and later analyzed using a Bio-Plex multiplex cytokine assay (Bio-Rad Laboratories, Hercules, CA, United States). CSF IL-10 levels were determined according to a standard curve generated for the specific target and expressed as picograms/milliliter (pg/mL). Samples were analyzed in triplicate.

Dolcetti E., Buttari F., Bruno A., Azzolini F., Gilio L., Di Caprio V., Lauritano G., Borrelli A., Galifi G., Furlan R., Finardi A., Musella A., Guadalupi L., Mandolesi G., Rovella V., Centonze D, & Stampanoni Bassi M. (2024). Low-contrast visual acuity test is associated with central inflammation and predicts disability development in newly diagnosed multiple sclerosis patients. Frontiers in Neurology, 15, 1326506.

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Multiplex Cytokine Profiling of Serum

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Blood serum was analyzed by the Bio-Plex multiplex cytokine assay (Bio-Rad Laboratories, Hercules, CA, USA) according to the manufacturer's instructions. The assay was read using a Luminex 100 (Austin, TX) and analyzed using a Bio-Plex Manager software. The mean concentration of cytokines (IL-4, IL-13, and LIF) in supernatants from OVA-stimulated cells over the unstimulated cells (background) was then calculated.

Hong S.H., Kim S.R., Choi H.S., Ku J.M., Seo H.S., Shin Y.C, & Ko S.G. (2014). Effects of Hyeonggaeyeongyo-Tang in Ovalbumin-Induced Allergic Rhinitis Model. Mediators of Inflammation, 2014, 418705.

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Investigating Cytokine Responses to CP4 Microvesicles

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To determine whether CP4 MVs could induce inflammation reactions in vitro, RAW264.7 cells were seeded in 24-well cell culture plates to 80% confluence. After washing once with DPBS, cells were exposed to 1 µg/ml, 5 µg/ml or 10 µg/ml MVs in fresh medium for 24 h at 37°C. Supernatants were collected then and the inflammation cytokines of both experimental and control (medium alone) were determined by Bio-Plex Multiplex Cytokine Assay (Bio-Rad). The serum samples from mice were used to determine the cytokine production in vivo.

Jiang Y., Kong Q., Roland K.L, & Curtiss R I.I.I. (2014). Membrane vesicles of Clostridium perfringens Type A strains induce innate and adaptive immunity. International journal of medical microbiology : IJMM, 304(0), 431-443.

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CSF Inflammatory Cytokine Profiling

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The CSF was collected by lumbar puncture (LP). Of note, 2 ml was used for the biochemical analyses, including total cell count and lactate levels. The presence of oligoclonal bands (OCB) was assessed. After LP, CSF was centrifuged and stored at −80°C. Pro-inflammatory and anti-inflammatory molecules were analyzed using a Bio-Plex multiplex cytokine assay (Bio-Rad Laboratories, Hercules, CA, USA). The CSF molecules examined were included as follows: interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12, IL-13, IL-15, IL-17, IL-1ra, granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), IP-10, interferon (IFN)-γ, tumor necrosis factor (TNF), monocyte chemoattractant protein 1 (MCP-1)/CCL2, regulated on activation, normal T cell expressed and secreted (RANTES), macrophage inflammatory protein 1-alpha (MIP-1α)/CCL3. All samples were analyzed in triplicate. Concentrations were expressed as picograms/milliliters.

Stampanoni Bassi M., Gilio L., Iezzi E., Moscatelli A., Pekmezovic T., Drulovic J., Furlan R., Finardi A., Mandolesi G., Musella A., Galifi G., Fantozzi R., Bellantonio P., Storto M., Centonze D, & Buttari F. (2021). Age at Disease Onset Associates With Oxidative Stress, Neuroinflammation, and Impaired Synaptic Plasticity in Relapsing-Remitting Multiple Sclerosis. Frontiers in Aging Neuroscience, 13, 694651.

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Cytokine Profiling in CSF

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CSF was centrifuged and immediately stored at −80°C until analyzed using a Bio-Plex multiplex cytokine assay (Bio-Rad Laboratories, Hercules, CA, USA) according to the manufacturer’s instructions. Concentrations of IL-1β (171-A11127; Bio-Rad Laboratories) were calculated according to a standard curve generated for each target and expressed in picograms per milliliter. When the cytokine concentrations were below the detection threshold, they were assumed to be 0 pg/ml.

Rossi S., Studer V., Motta C., Germani G., Macchiarulo G., Buttari F., Mancino R., Castelli M., De Chiara V., Weiss S., Martino G., Furlan R, & Centonze D. (2014). Cerebrospinal fluid detection of interleukin-1β in phase of remission predicts disease progression in multiple sclerosis. Journal of Neuroinflammation, 11, 32.

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CSF miR-142-3p and IL-1β Analysis

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CSF was collected at the time of diagnosis, during hospitalization, by lumbar puncture (LP) for medical purpose. No corticosteroids or disease modifying therapies were administered before LP. Cell-free CSF samples were stored at −80 °C and later analyzed [6 (link)]. Briefly, the CSF miR-142-3p levels were detected by quantitative real-time PCR (QIAGEN, Hilden, Germany) and normalized to miR-204-5p [6 (link)]. Samples were analyzed in duplicate. CSF concentrations of IL-1β were determined by using a Bio-Plex multiplex cytokine assay (Bio-Rad Laboratories, Hercules, CA, USA) and expressed in picograms/milliliter (pg/mL) according to the standard curve. Samples were analyzed in triplicate.

Dolcetti E., Musella A., Balletta S., Gilio L., Bruno A., Stampanoni Bassi M., Lauritano G., Buttari F., Fresegna D., Tartacca A., Mariani F., Palmerio F., Rovella V., Ferese R., Gambardella S., Giardina E., Finardi A., Furlan R., Mandolesi G., Centonze D, & De Vito F. (2024). Interaction between miR-142-3p and BDNF Val/Met Polymorphism Regulates Multiple Sclerosis Severity. International Journal of Molecular Sciences, 25(10), 5253.

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Cytokine Profiling in Cerebrospinal Fluid

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Immediately after LP, CSF samples were centrifuged and stored at −80°C. To analyze the levels of the main proinflammatory and anti-inflammatory molecules, a Bio-Plex multiplex cytokine assay (Bio-Rad Laboratories, Hercules, CA, USA) was used. The CSF levels of TNF and IL-6 were assessed. Concentrations were calculated according to a standard curve generated for the specific target and expressed as picograms/milliliter (pg/ml). All samples were analyzed in triplicate.

Stampanoni Bassi M., Gentile A., Iezzi E., Zagaglia S., Musella A., Simonelli I., Gilio L., Furlan R., Finardi A., Marfia G.A., Guadalupi L., Bullitta S., Mandolesi G., Centonze D, & Buttari F. (2019). Transient Receptor Potential Vanilloid 1 Modulates Central Inflammation in Multiple Sclerosis. Frontiers in Neurology, 10, 30.

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