The human inflammatory cytometric bead array (CBA) kit was purchased from BD Biosciences (San Jose, CA, USA). This kit was used to measure the levels of IL-1β, IL-6, IL-8, IL-10, IL-12p70 and TNFα, following the manufacturer’s instructions in supernatants collected from stimulated PBMC. Briefly, 50 µL of mixed beads were incubated with 50 µL of samples or standards and PE-labeled detecting antibodies at room temperature for 3 h. Following one wash of the beads with wash buffer, the beads were resuspended in 100 µL of buffer and analyzed by flow cytometry. Flow cytometry data were analyzed using FlowJo (Tree Star, V10.5.3). The concentrations of cytokines were calculated based on interpolation from a curve generated using the data from the kit’s standards. Cytokine induction was calculated using the following formula: cytokine concentration of stimulated PBMC/cytokine concentration of untreated PBMC.
Cba kit
Manufactured by BD
Sourced in United States, Germany, United Kingdom
The CBA kit is a laboratory equipment product from BD. It is designed for the quantitative measurement of cytokines and chemokines in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Fcap array software, by BD (8 mentions)
Facscalibur, by BD (6 mentions)
Facscalibur flow cytometer, by BD (5 mentions)
Cba software, by BD (5 mentions)
Facscanto 2, by BD (5 mentions)
Facscanto 2 flow cytometer, by BD (3 mentions)
Fcap array software version 3, by BD (3 mentions)
C57bl 6j b6 mice, by Jackson ImmunoResearch (2 mentions)
Brefeldin a, by BioLegend (2 mentions)
Concanavalin a (cona), by Merck Group (2 mentions)
Lsrii flow cytometer, by BD (2 mentions)
Diva software, by BD (2 mentions)
Legendplex software, by BioLegend (2 mentions)
Legendplex human inflammation panel cba, by BioLegend (2 mentions)
Recombinant mouse ifn γ, by Thermo Fisher Scientific (1 mentions)
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
Inomycin, by Merck Group (1 mentions)
Cck 8 kit, by Vazyme (1 mentions)
Microbeta trilux liquid scintillation counter, by PerkinElmer (1 mentions)
Cellquest pro, by BD (1 mentions)
111 protocols using cba kit
1
Cytokine Profiling of Stimulated PBMCs
2
Cytokine and Chemokine Quantification in Murine Immune Cells
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To detect IFN-γ, IL-2, IL-4, or IL-17A protein levels, 2 × 105 mononuclear cells or 105 CD4+ T cells from the indicated organs of pAAV- or pAAV-HBV1.2–injected mice were separated and cultured in 96-well U-bottomed plates in the presence of 2 µg/ml anti-CD3 plus 1 µg/ml anti-CD28 for 96 h. In some indicated cases, CD4+ T cells (105) were stimulated with 2 µg/ml HBcAg (ID LABS) or 5 µg/ml HBsAg (Hytest) in the presence of autologous irradiated splenocytes for 96 h. Supernatants were then collected, and cytokine levels were measured using their corresponding CBA kit (BD) according to the manufacturer’s instructions. To detect CXCL9, a total of 105 hepatocytes, LNPCs, F4/80+ Kupffer cells, or F4/80− LNPCs were cultured for 24 h; in some cases, the cells were cultured in the presence of 50 ng/ml recombinant mouse IFN-γ (PeproTech). Supernatants were also collected to measure CXCL9 protein levels using a CXCL9 CBA kit (BD). To measure the CXCL9 levels in liver, tissues were removed, weighed, and homogenized for CBA assay, as previously described (Hou et al., 2009 (link)).
3
Cytokine Analysis of Tumor Cell Co-culture
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Tumor cells (5 × 104) were co-cultured with the same amount of effector cells for 18 h and then centrifuged. The cell supernatant was collected and the amount of cytokines in the supernatant was analyzed by using the cytometric bead array (CBA) Kit (560484; BD Bioscience).
4
Adoptive Transfer of P14 Cells and LCMV Infection
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Six- to twelve-week old female C57BL/6j (B6) mice were purchased from Jackson laboratories. CD45.1+ or Thy1.1+ P14 transgenic mice that have TCR transgene specific for GP33 epitope were maintained in our animal facility. 1-2 × 104 P14 cells were adoptively transferred into B6 mice, followed by infection with LCMV Armstrong strain (2 × 105 PFU, intraperitoneally). For early activation experiments (Supplementary Fig. 6c, d ), P14 transgenic mice were directly infected with LCMV Armstrong strain (2 × 106 PFU, intravenously). For LCMV clone 13 strain infection, mice that received 2 × 103 P14 cells were intravenously infected with clone 13 (2 × 106 PFU). Viral titers in spleen were measured by plaque assay as described previously34 (link). Serum IFN-γ was measured by CBA kit (BD) according to manufacturer’s instruction. All animal experiments were approved by the Institutional Animal Care and Use Committee of Emory University.
5
Tumor-Infiltrating Lymphocyte Activation
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Spleens and lymph nodes of tumor-burdened mice were ground through 70 μm filter strainers and washed with RPMI 1640. Red blood cell lysis was performed according to the protocol described above. Lymphocytes (4 × 105) were incubated in 96-well plates in the absence or presence of OT-I or OT-II peptide at 37 °C and 5% CO2. IFN-γ and TNF-α in the supernatant were measured using a CBA kit (BD Biosciences, USA).
6
Cytokine Profiling in Murine Arthritis
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Cell culture supernatants were collected from cell culture well plates at the indicated time points and sera were collected from the different groups of CIA mice. IFN-γ, TNF, IL-17a, IL-2, IL-4, IL-6, and IL-10 concentrations were measured using a CBA kit (BD Biosciences). T cells were isolated from spleens and draining lymph nodes of the CIA mice at day 63 after CII immunization. The cells were then stimulated with PMA (Sigma, P8139), inomycin (Sigma, I0634) and Brefeldin A (Biolegend, 10 μg/mL) for 4 h, and intracellular IFN-γ and IL-17a expression were analyzed by flow cytometry.
7
Immunomodulatory Cytokine Profiling
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The lymphocyte proliferation was determined every day from the first to the fifth day using a CCK8 kit (Vazyme Biotech, China). The optical absorbance was measured at 450 nm by a microplate reader. The cytokines in the culture medium were analyzed using a CBA kit (BD Bioscience, United States); the assay was carried out according to the manufacturer’s instructions. Data were acquired with a BD FACS Calibur flow cytometer and analyzed with the FCAP array software. IL-2, IL-4, IL-6, IFN-g, TNF-a, IL-17, and IL-10 were recorded and compared.
8
Cytokine profiling of influenza-specific T cells
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Splenocytes (one million) from mice groups at 10 days post-dose-2 were cultured and stimulated with 1 µg HA + 1 μg NP + 5 μg M2e at 37°C for 4/24/72 h. The culture supernatants were tested for Th1 cytokines (IFN-γ, IL-2, and TNF) and Th2 cytokines (IL-4, IL-6, IL-10, and IL-5) by using CBA kit (BD Biosciences, USA) as described previously (30 (link)). As positive control, splenocytes stimulated with 2.5 µg/well concanavalin A (Sigma, USA) and as negative control, unstimulated cells were used. Cells collected after 4 and 24 h of stimulation were used for surface staining and gene expression analysis by TLDA.
9
T Cell Costimulation Assay with FGL1 and LAG3
10
Cytokine Response to CMFO Restimulation
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10 weeks after immunization, splenocytes were prepared and counted from each immunized mice in different groups. 5 × 106 cells were seeded in each well of a 24-well microtiter plate in triplicate and restimulated with 10 μg/mL CMFO for 72 h as previously described [15 (link), 16 (link)]. 72 hours later, supernatant for each well was collected and a cytometric bead array (CBA) kit (BD Biosciences, USA) was used to detect the concentration of CMFO-specific Th1/Th2/Th17 cytokines containing IFN-γ, IL-2, TNF-α, IL-4, IL-6, IL-10, and IL-17 [16 (link)]. The results were expressed as the mean ± SEM (pg/mL) for each group (n = 6).
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