Cytometric bead array human th1 th2 th17 cytokine kit

Manufactured by BD

Sourced in United States

The Cytometric Bead Array (CBA) Human Th1/Th2/Th17 Cytokine Kit is a laboratory tool designed to measure the concentration of multiple cytokines in a single sample. It utilizes a flow cytometry-based platform to simultaneously quantify the levels of specific Th1, Th2, and Th17 cytokines.

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Lab products found in correlation

Facscalibur flow cytometer, by BD (4 mentions) Facscanto 2 flow cytometer, by BD (3 mentions) Fcap array software, by BD (3 mentions) Fcap array software v3, by BD (2 mentions) Rpmi medium, by Thermo Fisher Scientific (2 mentions) Aim 5, by Thermo Fisher Scientific (2 mentions) Ab serum, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Roche (2 mentions) Ultra leaf anti cd3 ab okt3, by BioLegend (2 mentions) Ultra leaf anti cd28 ab cd28, by BioLegend (2 mentions) Lcki inhibitor, by Merck Group (2 mentions) Penicillin, by Roche (2 mentions) Anti cd3 cd28 beads, by Thermo Fisher Scientific (2 mentions) Anti cd3 cd28 beads, by BD (2 mentions) Accuri c6 flow cytometer, by BD (2 mentions) Accuri c6 sampler, by BD (2 mentions) Facsuite v 1, by BD (1 mentions) Facsverse, by BD (1 mentions) Flow cytometry, by BD (1 mentions) Sandwich elisa, by R&D Systems (1 mentions)

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Cytometric bead array (430 citations) Cytokine Bead Array (59 citations)

37 protocols using cytometric bead array human th1 th2 th17 cytokine kit

Cytokine Release Profiling by Flow Cytometry

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Concentration of several cytokines, released during biopeptide treatment, was determined using flow cytometry (Becton Dickinson, NJ, USA) [65 (link),66 (link)].
The release of cytokines (IL-2, IL-4, IL-6, IL-10, TNF, INF-γ, IL17-A) in supernatants was measured by flow cytometry using the commercial Cytometric Bead Array (CBA) Human Th1/Th2/Th17 Cytokine Kit (BD Bioscience catalogue 560484), following the manufacturer’s instructions. Samples acquisition was performed on FACSVerse (BD Biosciences) with FACSuite v 1.0.6.5230 (BD Biosciences) and data analysis by FCAP Array Software v3.0 (BD Bioscience) as suggested.

Avolio F., Martinotti S., Khavinson V.K., Esposito J.E., Giambuzzi G., Marino A., Mironova E., Pulcini R., Robuffo I., Bologna G., Simeone P., Lanuti P., Guarnieri S., Trofimova S., Procopio A.D, & Toniato E. (2022). Peptides Regulating Proliferative Activity and Inflammatory Pathways in the Monocyte/Macrophage THP-1 Cell Line. International Journal of Molecular Sciences, 23(7), 3607.

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Quantification of Serum Cytokine Levels

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The serum was isolated from 4 ml of blood obtained in tubes without additives from each individual and stored at −80°C until analysis. The quantitative determination of inflammatory cytokines in serum was performed using the Cytometric Bead Array (CBA) Human Th1/Th2/Th17 Cytokine kit (BD Biosciences; San Diego, California, USA). This kit allows quantitatively measure interleukin (IL) 2, IL-4, IL-6, IL-10, Tumor Necrosis Factor (TNF) α, Interferon (IFN) γ, and IL-17A protein levels in a single sample. The fluorescence produced by CBA beads was measured on a FACS Canto II Flow Cytometer (BD Biosciences) and analyzed using FCAP array software (Soft Flow Inc; New Brighton, MN, USA). Detection limits were 2.6 pg/ml for IL-2, 4.9 pg/ml for IL-4, 2.4 pg/ml for IL-6, 4.5 pg/ml for IL-10, 3.8 pg/ml for TNF-α, 3.7 pg/ml for IFN-γ, and 18.9 pg/ml for IL-17A.

Álvarez-Rodríguez L., Martínez-Taboada V., Calvo-Alén J., Beares I., Villa I, & López-Hoyos M. (2019). Altered Th17/Treg Ratio in Peripheral Blood of Systemic Lupus Erythematosus but Not Primary Antiphospholipid Syndrome. Frontiers in Immunology, 10, 391.

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Cytokine Profiling for CRS and Neurotoxicity

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The BD^™ Cytometric Bead Array (CBA) Human Th1/Th2/Th17 Cytokine Kit (San Jose, CA, USA) was used to analyze serum concentrations of released cytokines, including interleukin (IL)-2, IL-4, IL-6, IL-10, IL-17A, tumor necrosis factor α (TNF-α), and interferon γ (IFN-γ). Cytokine release syndrome (CRS) was graded according to the American Society for Transplantation and Cellular Therapy (ASTCT) grading system [23 ]. Neurologic toxicity was graded in accordance with the Common Terminology Criteria for Adverse Events (CTCAE) 4.03 [24 ].

Li W., Ding L., Shi W., Wan X., Yang X., Yang J., Wang T., Song L., Wang X., Ma Y., Luo C., Tang J., Gu L., Chen J., Lu J., Tang Y, & Li B. (2023). Safety and efficacy of co-administration of CD19 and CD22 CAR-T cells in children with B-ALL relapse after CD19 CAR-T therapy. Journal of Translational Medicine, 21, 213.

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Cytokine Profiling in HPV Infection

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The levels of cytokines were determined in the serum and ECCs from 26 patients positive for HR-HPV DNA and 18 patients negative for HPV DNA. The concentrations of IFN-γ, TNF-α, IL-10, IL-6, IL-4, and IL-2 cytokines were determined using a Cytometric Bead Array (CBA) Human Th1/Th2/Th17 Cytokine Kit (BD Biosciences, San Jose, CA, USA). Standards, beads, samples, and protocols for the flow cytometer setup and data acquisition were performed according to the manufacturer’s instructions. Samples were evaluated using a FACS Canto II Flow Cytometer (BD Biosciences, San Jose, CA, USA) and analyzed with the FCAP Array software V. 3.0 (Becton Dickinson, Franklin Lakes, NJ, USA). The results were based on standard concentration curves and expressed as picograms per milliliter (pg/mL).

Bonin-Jacob C.M., Almeida-Lugo L.Z., Puga M.A., Machado A.P., Padovani C.T., Noceti M.C., Ferreira A.M., Fernandes C.E., Resende J.C., Bovo A.C, & Tozetti I.A. (2021). IL-6 and IL-10 in the serum and exfoliated cervical cells of patients infected with high-risk human papillomavirus. PLoS ONE, 16(3), e0248639.

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Cytokine Profile Analysis in Serum

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The blood was sampled at the follow-up time points. The protein levels of interleukins (IL-2, IL-4, IL-6, IL-10 and IL-17A), interferon (IFN)-γ and tumor necrosis factor (TNF) were evaluated using the Cytometric Bead Array (CBA) Human Th1/Th2/Th17 Cytokine Kit (BD Biosciences, San Jose, CA, USA) and flow cytometry analysis. All assays were carried out according to the manufacturer’s protocol. Flow cytometry was used to detect the cytokine levels in serum samples. The serum data were expressed in picograms per milliliter.

Ying Z., He T., Jin S., Wang X., Zheng W., Lin N., Tu M., Xie Y., Ping L., Liu W., Deng L., Ding Y., Hu X., Bu B., Lu X., Song Y, & Zhu J. (2022). A durable 4-1BB-based CD19 CAR-T cell for treatment of relapsed or refractory non-Hodgkin lymphoma. Chinese Journal of Cancer Research, 34(1), 53-62.

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CD8+ T cell activation and IFNγ analysis

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Isolated CD8⁺ T cells from PBMCs (see above) were activated in 96-well plates coated with 5mg/ml plate bound Ultra-LEAF™ anti-CD3 Ab (OKT3, Biolegend) and 2mg/ml plate bound Ultra-LEAF™ anti-CD28 Ab (CD28.2, Biolegend) at a density of 1 × 10⁵ cells per well, in a 1:1 mix of Aim 5 (Gibco) and RPMI medium (Gibco), supplemented with 10% AB serum (Life Technologies), 100 U/ml penicillin (Roche), 100 mg/ml streptomycin (Roche). After 2 h, cells were either left untreated or were treated with 5 nM LCKi inhibitor (Merck Milipore) for the indicated time periods. Cells were subsequently washed 3 times with medium to remove already secreted IFNγ, and cells were then cultured for 3h in 40 ml fresh control medium or fresh medium containing 5 nM LCK inhibitor. Subsequently, supernatants were collected and IFNγ concentrations were determined using the BD™ Cytometric Bead Array (CBA) Human Th1/Th2/Th17 Cytokine Kit, according to the manufacturer’s protocol.

Hoekstra M.E., Bornes L., Dijkgraaf F.E., Philips D., Pardieck I.N., Toebes M., Thommen D.S., van Rheenen J, & Schumacher T.N. (2020). Long-distance modulation of bystander tumor cells by CD8+ T cell-secreted IFNγ. Nature cancer, 1(3), 291-301.

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Quantifying Th1/Th2/Th17 Cytokines

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Cytokine concentrations were estimated in culture supernatants using the Cytometric Bead Array (CBA) Human Th1/Th2/Th17 Cytokine Kit (BD) or sandwich ELISAs (R&D), as indicated.

Canavan J.B., Scottà C., Vossenkämper A., Goldberg R., Elder M.J., Shoval I., Marks E., Stolarczyk E., Lo J.W., Powell N., Fazekasova H., Irving P.M., Sanderson J.D., Howard J.K., Yagel S., Afzali B., MacDonald T.T., Hernandez-Fuentes M.P., Shpigel N.Y., Lombardi G, & Lord G.M. (2015). Developing in vitro expanded CD45RA+ regulatory T cells as an adoptive cell therapy for Crohn's disease. Gut, 65(4), 584-594.

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Cytokine Quantification Using CBA Kit

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Cytokine quantification was performed using a BD^™ Cytometric Bead Array (CBA) Human Th1/Th2/Th17 cytokine kit using supernatants from the ex-vivo IFNγ ELISpot. 35μL of the pooled peptide triplicate supernatant from the DMSO, Fa, Fb, M and N wells were mixed with 5μL aliquot of each cytokine capture bead (human IL2, IL4, IL6, IL10, TNF, IFNγ and IL17A) and 35μL detection reagent (phycoerythrin(PE)-conjugated antibody) for 3 hours at room temperature and protected from light. 800μL of wash buffer was then added and each sample centrifuged at 200g for 5 minutes. The supernatant was discarded and the bead pellet was re-suspended in 200μL wash buffer. Cytokine detection was performed using an LSRII FACS machine (BD), BD FACSDiva software (version 6.0 for Windows) and FlowJo software (version X0.7 for Mac).

Green C., Scarselli E., Sande C., Thompson A., de Lara C., Taylor K., Haworth K., Del Sorbo M., Angus B., Siani L., Di Marco S., Traboni C., Folgori A., Colloca S., Capone S., Vitelli A., Cortese R., Klenerman P., Nicosia A, & Pollard A. (2015). Chimpanzee adenovirus and MVA-vectored respiratory syncytial virus vaccine is safe and expands humoral and cellular immunity in adults. Science translational medicine, 7(300), 300ra126.

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Cytokine Release Quantification

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Collected the supernatant during cytotoxicity assays to measure released cytokines using Cytometric Bead Array (CBA) Human Th1/Th2/Th17 Cytokine Kit (BD, Franklin Lakes, NJ). The data were analyzed using Flowjo10.8.1 software.

Xiong Q., Wang H., Shen Q., Wang Y., Yuan X., Lin G, & Jiang P. (2024). The development of chimeric antigen receptor T-cells against CD70 for renal cell carcinoma treatment. Journal of Translational Medicine, 22, 368.

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CD8+ T cell activation and IFNγ analysis

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