CD26 2/3G CAR-T-cells and CD8 2/3G control cells were placed onto a 96-well culture plate (1 × 105 cells/well) as the effector cells and were co-cultured overnight with CD26-Jurkat or HSB2 cells as the target cells. For positive controls of the activated effector cells, effector cells were incubated with both 10 ng/mL of phorbol-12-myristate 13-acetate (PMA) (FUJIFILM Wako, Osaka, Japan) and 500 ng/mL of ionomycin (Merck, Darmstadt, Germany) for the same period without the target cells. CD69 expression on effector cells, which were selected from the target cells by gating on marker fluorescence GFP, was then analyzed by flow cytometry using an APC anti-human CD69 antibody, clone FN50 (BioLegend). Furthermore, concentrations of interferon gamma (IFN-γ) and granzyme B in the culture supernatants secreted by the effector cells were measured by the Enzyme-Linked Immuno Sorbent Assay (ELISA) method using a Human IFN-gamma DuoSet ELISA (R&D Systems, Minneapolis, MN) and a Human Granzyme B DuoSet ELISA (R&D Systems). The concentrations of tumor necrosis factor-alpha (TNFα) and inerleukin-8 (IL-8) were also measured by flow cytometry using a BDTM Cytometric Bead Array (CBA) Kit (BD Biosciences) according to the manufacturer’s instructions.
Cytometric bead array cba kit
Manufactured by BD
Sourced in United States
The BD Cytometric Bead Array (CBA) kit is a multiplex assay designed to quantify multiple analytes in a single sample. The kit utilizes fluorescent-coded beads that are coated with capture antibodies specific to the target analytes. The sample is incubated with the beads, and the bound analytes are detected using a flow cytometer.
Automatically generated - may contain errors
Lab products found in correlation
Fcap array software, by BD (3 mentions)
Ionomycin, by Merck Group (1 mentions)
Phorbol myristate acetate (pma), by Fujifilm (1 mentions)
Human granzyme b duoset elisa, by R&D Systems (1 mentions)
Human ifn gamma duoset elisa, by R&D Systems (1 mentions)
Opteia, by BD (1 mentions)
Facscanto 2 flow cytometer, by BD (1 mentions)
Lsrfortessa, by BD (1 mentions)
Anti mouse cd3 clone 17a2, by BioLegend (1 mentions)
Quantikine elisa, by R&D Systems (1 mentions)
Facsverse, by BD (1 mentions)
Fluostar omega microplate reader, by BMG Labtech (1 mentions)
24 well tissue culture plate, by Corning (1 mentions)
15 protocols using cytometric bead array cba kit
1
Evaluating CD26 CAR-T Cell Cytotoxicity
2
Cytokine Profiling in J774A.1 Cells
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Cytokine production in J774A.1 cells was quantified using the mouse inflammation BDTM Cytometric Bead Array (CBA) kit (50TST, BD Biosciences, San Diego, CA, USA). Flow cytometry was performed using The BD LSR Fortessa™ cell analyzer. Results were normalized to the total protein concentration. Assays were performed according to the manufacturers´ instructions. The measured cytokines were: Interleukin-6 (IL-6), Interleukin-10 (IL-10), Monocyte Chemoattractant Protein -1 (MCP-1) and Tumor Necrosis Factor-α (TNF-α).
3
Pep-H Modulates Host Immune Response
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Apart from direct antimycobacterial activity, effect of Pep-H on host immune system was also evaluated. To measure cytokines levels in the respective culture supernatants, BD Cytometric Bead Array (CBA) kit was used. IL-8 and MCP-1 concentrations were measured using anti human IL-8 and MCP-1 reagents (OptEIATM, BD Pharmingen, USA). Nitrite levels were determined by the Griess assay54 (link).
4
Quantification of IL-2 Secretion in T Cells
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IL-2 levels were measured with BD Cytometric Bead Array (CBA) kit. Briefly, 50 K TCM CD8 T cells were stimulated with anti-CD3/CD28 beads in a 96-well round-bottom plate. After 18 h, beads were removed by plate magnet then cultured alone for 7 days then supernatants were collected and stored at −80 Celsius. At the day of the experiment, 50 μL of thawed supernatant was added to 50 μl of IL-2 capture beads (beads coated with anti-IL-2 antibody) then incubated for 2 h at room temperature in dark. Subsequently, 50 μL of phycoerythrin (PE) detection reagent was added then incubated for 1 h at room temperature in the dark. The samples were washed with 1 mL of wash buffer and centrifuged for 5 min at 200 g. After discarding the supernatant, the bead pellet was resuspended in 100 μL buffer. Measurements were performed on the Flow Cytometer (BD Fortessa) and then analyzed by FCAP ArrayTM Software (BD Bioscience). IL-2 concentration was measured by its fluorescent intensity. Calibrated standard curve was generated using the IL-2 standard serial dilutions.
5
Quantifying Interleukin-8 in Seminal Plasma
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Interleukin-8 was measured in culture medium supplemented with seminal plasma using a BD Cytometric Bead Array (CBA) kit (BD Bioscience, San Jose, CA). The cytokine was analyzed following the manufacturer’s instructions using BD FACS Canto II Flow Cytometer and FCAP ArrayTM Software (BD Bioscience). GraphPad Prism (GraphPad Software, Inc., La Jolla, CA) was used in the statistical analyses. Statistical significance was determined at 95% (p<0.05).
6
Effects of 1-methyl-tryptophan on cytokines
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The TC-1 cells were seeded in 6-well cell dishes (1,5x105/well) and cultured in RPMI 1640 medium supplemented with 10% of fetal bovine serum (FBS) (R10) until reaching 50-60% confluency. Next, cells were treated with a fresh R10 medium containing 1mM of 1-methyl-D-tryptophan (D-1MT), 1-methyl-L-tryptophan (L-1MT), or 1-methyl-DL-tryptophan (DL-1MT) and incubated for 24 hours at 37°C and 5% CO2. In the control group, only R10 medium was added. Cell culture supernatant was collected for cytokine measurement by BD™ Cytometric Bead Array (CBA) kit (#560485, BD Biosciences). Stained samples were acquired by LSR Fortessa™ (BD Biosciences) flow cytometer and data were analyzed using FlowJo software (TreeStar).
7
Lymphocyte Profiles and Cytokine Responses in Influenza B Infection
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Nasopharyngeal swabs were obtained within 24 hours after admission, and influenza B virus RNA was detected in exfoliated cells by polymerase chain reaction (PCR) or nucleic acid hybridization. Venous blood samples were collected within 24 hours after admission, flow cytometry analysis was used to detect the absolute count and percentage of the total T cell (CD3+), CD4+ T cell (CD3+CD4+), CD8+ T cell (CD3+CD8+), NK cell (CD16+CD56+), the total B cell (CD19+) on the surface of lymphocytes, and the ratio of CD4+/CD8+ (Th/Ts). We used BD™ Cytometric Bead Array (CBA)Kit based on flow cytometry detected cytokines (IL-2, IL-4, IL-6, IL-10, IL-17, INF-γ, IL-12p70, IL-1β, TNF-α) in peripheral blood.
8
Cytokine Release Analysis of MSCs
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The committed MSCs are seeded into a 96-well plate at a density is 1 × 104 cells/cm2 and incubated overnight. The cell medium was collected after treating with different concentrations of NPs for analyzing IL-1α, IFN-γ and TNF-α by BD cytometric bead array (CBA) kit (BD Biosciences, NJ, USA). The kit was performed according to the manufacturer’s instructions and analyzed by flow cytometry.
9
Evaluating VISTA Protein Modulation of CD8+ T Cells
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CD8+ T cells were isolated as described above, and a density of 1 × 105 cells/well were plated into the 96-well flat-bottom plates coated with anti-mouse CD3 (clone 17A2, Biolegend) at 2.5 μg/mL together with 5 μg/mL of murine VISTA-ECD protein. The compound 6809-0223 diluted into indicated concentrations was added to the culture medium. The cytokine levels in the cell supernatant collected at 48 h were analyzed using a BD Cytometric Bead Array (CBA) kit according to the manufacturer’s instructions.
10
Cytokine Profiling in CD Patients
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The whole peripheral blood samples of CD patients and healthy people were centrifugated at 2000 rpm for 5 min to collect the serums. The cytokines (IL-2, IL-4, IL-6, IL-10, TNF-α, IFN-γ and IL-17A) in these serums were detected by using BD™ Cytometric Bead Array (CBA) kit according to the manufacturer's instruction.
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