Purified anti-CD3 and anti-CD28 (BD Pharmingen) were diluted to a concentration of 5 μg/mL in PBS and coated onto 96-well EIA/RIA plates. Unfractionated splenocytes were resuspended in complete media (CM), consisting of RPMI 1640 (VWR) supplemented with 5% FBS, 100 U/mL penicillin, 100 µg/mL streptomycin, 2 mM L-glutamine, and 0.05 mM β-mercaptoethanol. The cell suspension was applied to antibody-coated plates at a seeding density of 2 × 105 cells per well. Cultures were harvested after incubating at 37 oC for 24 h. For each sample, culture supernatants were pooled from three replicate wells. Cytokine levels in supernatants were assayed by flow cytometry using the BD Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 Cytokine Kit according to the manufacturer’s instructions. Samples were run on a BD LSRFortessa HTS flow cytometer and analyzed using BD FACSDiva software.
Cytometric bead array cba mouse th1 th2 th17 cytokine kit
Manufactured by BD
Sourced in United States
The BD Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 Cytokine Kit is a multiplex immunoassay system designed to measure the concentrations of seven mouse cytokines (IFN-γ, IL-2, IL-4, IL-5, IL-6, IL-10, and IL-17A) simultaneously in a single sample.
Automatically generated - may contain errors
Lab products found in correlation
Facscanto 2, by BD (5 mentions)
Fcap array software, by BD (3 mentions)
96 well plate, by Corning (3 mentions)
Facscalibur flow cytometer, by BD (3 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (2 mentions)
Concanavalin a (cona), by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (2 mentions)
Fcap array software version 3, by BD (2 mentions)
Rpmi 1640, by Avantor (1 mentions)
Lsrfortessa hts flow cytometer, by BD (1 mentions)
Purified anti cd3 and anti cd28, by BD (1 mentions)
Fortessa flow cytometer, by BD (1 mentions)
Human aβ42 elisa kit, by Thermo Fisher Scientific (1 mentions)
Lsrfortessa, by BD (1 mentions)
Facs calibur flow cytomyter, by BD (1 mentions)
Miltefosine, by Merck Group (1 mentions)
Platinum sybr green, by Thermo Fisher Scientific (1 mentions)
Permount mounting medium, by Thermo Fisher Scientific (1 mentions)
Purelink genomic dna mini kit, by Thermo Fisher Scientific (1 mentions)
34 protocols using cytometric bead array cba mouse th1 th2 th17 cytokine kit
1
T-cell Activation and Cytokine Profiling
2
Cytokine Profiling in Blood Plasma
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The withdrawn blood was centrifuged at 5000 rpm for 10 minutes to separate the plasma and peripheral blood mononuclear cells. Cytokine levels in blood plasma were then determined using a BD Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 Cytokine Kit (BD Biosciences, San Jose, CA) according to manufacturer’s instructions. Cytokines IL-2, IL-4, IL-6, IFN-γ, TNF-α, IL-17A and IL-10 were quantified using FACSCanto II (BD Biosciences) and analysed using FCAP array software (BD Biosciences). TGF-β cytokine in blood plasma was quantified using Human/Mouse TGF-β1 sandwich ELISA (eBiosciences, San Diego, CA) based on manufacturer’s instructions.
3
Multiplex Cytokine Profiling in Mice
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For detection of various cytokines, plasma samples were collected from mice or cells at indicated time after Con A injection and immediately frozen at −70 °C and stored until use. BD Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 Cytokine Kit (for GM-CSF, IL-1b, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IFN-γ and tumor necrosis factor (TNF)-α) was purchased from BD Biosciences. All procedures were performed according to the manufacturer’s protocol.
4
Multiplex Cytokine Profiling in Serum
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Detection of cytokines was carried out using the commercially available BD cytometric bead array (CBA) Mouse Th1/TH2/TH17 cytokine kit (BD, San Jose, CA, USA) as established previously with some minor adaptions (2 (link)). In brief, 10 μL of serum sample was used in 96 well plates and reagent, containing mouse cytokine capture beads with distinct fluorescent intensities, was proportionally adjusted to suit sample size. Samples were measured using a BD Fortessa Flow Cytometer (BD, San Jose, CA, USA) and all sample analysis and data were collected using FlowJo (FlowJo, OR, USA) flow cytometry software.
5
Quantitative Analysis of Alzheimer's and Inflammation
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For quantitative analysis of Aβ and inflammatory factors, frozen tissues of APP/PS1 mice treated with PBS, MSNs, MSNs-Bi, and Bifidobacterium were homogenized in liquid nitrogen and extracted using TBS solution. The homogenized supernatant was then centrifuged at 10,000 g for 30 min at 4 °C to obtain TBS-soluble proteins. Following this, Aβ levels in the mouse brain and intestine were quantified using the Aβ42 human ELISA kit (Catalog #KHB3441, Invitrogen). The BD Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 Cytokine kit (Catalog #560485, BD Biosciences) was used to quantify the levels of inflammatory factors in the blood.
6
Cytokine Profiling of Activated CD4+ T Cells
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CD4+ T cell from the spleens of the mice were incubated at a density of 2 × 105 per well in a 96-well plate. Cells were stimulated with Con A at various concentrations (1, 2.5, or 5 μg/ml) for up to 48 h. Supernatants (100 μl) were taken and immediately frozen at −70 °C and stored until detection with BD Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 Cytokine Kit (for GM-CSF, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IFN-γ and tumor necrosis factor (TNF) α).
7
Splenocyte Cytokine Profiling Protocol
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Two days before challenge, four mice from each group were randomly sacrificed, and cell suspensions were prepared under aseptic conditions by grinding the spleens and filtering through 200-mesh screens. The splenocytes from each mouse were cultured in triplicate (cell density: 5 × 105 cells per well) in 96-well plates (Corning; NY, USA), incubated in RPMI 1640 medium (Hyclone; South Lagan, USA) supplemented with 10% fetal calf serum (Gibco; Grand Island, USA), and stimulated with rTPI (10 μg/ml), ConA (Sigma-Aldrich; St. Louis, USA, 10 μg/ml), or medium alone (mock) at 37 °C with 5% CO2 for 72 h. The supernatants were collected, and cytokine levels were measured using a BD Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 Cytokine Kit, according to the manufacturer’s protocols.
8
Dendritic Cells Activation Assay
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DCs were seeded in a 6-well plate at a density of 1 × 106 cells per well. PBS, OVA, OVA@LP, OVA@CD11c-ILP with OVA concentration of 2 μg/mL and LPS (100 ng/mL) were added to the well and incubated for 24 h at 37 °C with 5% CO2. Then, cells and supernatant were collected separately. Cells were washed twice with PBS and resuspended in 100 μL buffers. Cells were labeled with different monoclonal antibodies (CD40, CD80, CD86, Iab) and then washed twice with cold PBS. Expression levels of DCs surface molecules, such as CD40, CD80, CD86 and Iab, were determined by flow cytometry. TNF-α in supernatant was examined by using the BD Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 Cytokine Kit.
9
Cytokine Profiling in Mouse Th Cells
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Serum cytokine levels of TNF‐α, IL‐10 and IL‐6 were measured using the BD Cytometric bead array (CBA) Mouse Th1/Th2/Th17 Cytokine Kit (BD Biosciences) according to the manufacturer's instructions. The LSR‐Fortessa (BD Biosciences) and FlowJo (Tree Star) were used for acquisition/analysis.
10
Cytokine Quantification in Mice
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Mouse IFN-γ, TNF, and IL-17A were measured using the BD cytometric bead array (CBA) Mouse Th1/Th2/Th17 Cytokine Kit (Cat#: BD 560485, San Diego, CA) in compliance with the manufacturer’s instructions.
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