The proinflammatory cytokines interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor α (TNFα) were measured in plasma obtained from the mice at sacrifice. Bead-based immunoassays BD Cytometric Bead Array (CBA), from BD Biosciences, San Jose, CA, USA, were used according to the manufacturer's instructions. Data were collected on a BD LSRII flow cytometer (BD Biosciences) and analysed by use of FCAP Array software 3.0 (BD Biosciences). All samples were diluted 1 : 2 in Assay Diluent included in the CBA kit. The limits of detection were 1.9, 1.4 and 2.8 pg/mL, for IL-1β, IL-6 and TNFα, respectively.
Fcap array software 3
Manufactured by BD
Sourced in United States
FCAP Array software 3.0 is a data analysis software for flow cytometry applications. It provides tools for visualizing and analyzing data from flow cytometry experiments.
Automatically generated - may contain errors
Lab products found in correlation
Cytometric bead array cba, by BD (1 mentions)
Lsr 2 flow cytometer, by BD (1 mentions)
Infinite m200 reader, by Tecan (1 mentions)
Facscalibur, by BD (1 mentions)
Lsrfortessa cell analyzer, by BD (1 mentions)
Cytometric bead array, by BD (1 mentions)
Facscanto 2 system, by BD (1 mentions)
Mouse inflammation kit, by BD (1 mentions)
Lsrfortessa flow cytometer, by BD (1 mentions)
Tri reagent, by Merck Group (1 mentions)
Quantitect reverse transcription kit, by Qiagen (1 mentions)
Facsdiva software, by BD (1 mentions)
Faststart sybr green master mix, by Roche (1 mentions)
Cytometric bead array mouse th1 th2 th17, by BD (1 mentions)
Facscanto 2 flow cytometer, by BD (1 mentions)
8 protocols using fcap array software 3
1
Measuring Inflammatory Cytokines in Mice
2
Cytokine Profiling of TGF-β1 Secretion
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Secretion of human TGF-β1 (BMS249, eBioscience) was analyzed by ELISA, used according to the manufacturer’s protocol. Samples were measured in duplicates with a Microplate Reader (Tecan Reader infinite® M200, Tecan, Crailsheim, Germany). IL-1ß, IL-6, IL-8, IL-10, IL-12 and TNF-α in cell culture supernatant were characterized by human Inflammatory Cytokine CBA (551811, BD Biosciences, Heidelberg, Germany) according to manufacturer’s protocol. Analysis was performed with FACS Calibur (BD Biosciences) and data was processed using FCAP Array Software 3.0 (BD Biosciences).
3
Quantification of Inflammatory Cytokines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The levels of TNF-α, IL-6, IL-10, MCP-1, IFN-γ, and IL-12p70 in blood were measured using a Mouse Inflammation CBA Kit (Becton Dickinson). Briefly, sera were diluted 10-fold in Assay Diluent and 50 μl of diluted sample was incubated with 50 μl of mixed Mouse Inflammation Capture Beads and 50 μl of Mouse Inflammation PE Detection Reagent for 2 hrs. After washing the mixture once in Wash Buffer, the intensities of the signals on beads were measured by flow cytometry, and the results were analyzed using FCAP Array software 3.0 (BD). All steps were performed at room temperature. Cytokine concentrations were expressed as pg/ml.
4
Cytokine Profiling of MHV-infected Macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
C57BL/6 BMDMs were infected with wild-type-, DUBmut-, or EndoUmut-MHV at an MOI of 1. At 24 h p.i., supernatants were collected and centrifuged at 1,000 × g for 5 min at 4°C to remove cellular debris. CBA staining was performed according to the manufacturer’s instructions (CBA Flex Set mouse soluble proteins, catalog no. 558266; BD Biosciences). Bead fluorescence was measured with an LSR Fortessa cell analyzer (BD Biosciences), and data were analyzed using FCAP Array software 3.0 (BD Biosciences).
5
Cytokine Profiling of T-Cell Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Supernatants of the IFN-γ ELISPOT assay and CD107a assay were collected and interleukine-2 (IL-2) and tumor necrosis factor (TNF) were measured using a Cytometric Bead Array (BD Bioscience) according to the manufacturer's instructions. Samples were analyzed using a FACSCanto II system and the FCAP Array Software 3.0 (BD Bioscience).
6
Evaluating Antiviral and Inflammatory Responses to AAV
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To detect IFN-β, OAS-1, and IFIT-1 transcript concentrations, livers from mice were harvested 6 hr after AAV injection and RNA isolated using TRI Reagent (Sigma). 1 μg of RNA was used for reverse transcription using a QuantiTect Reverse Transcription Kit (QIAGEN). Resultant cDNA was then used for SYBR green-based real-time qPCR using FastStart SYBR Green Master Mix (Roche Diagnostics).
To assess induction of release of inflammatory cytokines by AAVs, serum concentrations of IL-6, IL-10, MCP-1, IFN-γ, TNF-α, and IL-12p70 were measured. Assays using a mouse inflammation kit (BD Biosciences) were carried out on serum taken immediately before and 6 hr after scAAV administration to groups of mice, which each comprised four animals. CBA reactions were run on a Fortessa LSR flow cytometer using the BD FACSDiva software. Data were analyzed using FCAP Array software 3.0 (BD Biosciences).
To assess induction of release of inflammatory cytokines by AAVs, serum concentrations of IL-6, IL-10, MCP-1, IFN-γ, TNF-α, and IL-12p70 were measured. Assays using a mouse inflammation kit (BD Biosciences) were carried out on serum taken immediately before and 6 hr after scAAV administration to groups of mice, which each comprised four animals. CBA reactions were run on a Fortessa LSR flow cytometer using the BD FACSDiva software. Data were analyzed using FCAP Array software 3.0 (BD Biosciences).
7
Cytokine Profiling by CBA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cytokines in the serum samples were measured with Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 (BD Biosciences) according to the manufacturer’s instructions. The data were collected on a FACSCANTO II flow cytometer (BD Bioscience) and analysed by FCAP Array Software 3.0 (BD Bioscience).
8
Profiling Cytokines in Activated T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To determine the cytokine profiles of stimulated T cells, we use co-cultures with allogeneic Mo-DCs primed with adenoviral-transduced SK-MEL-147 cells. Supernatants were collected after 3 days of cultures and analyzed by CBA. Analysis of IL-2, IL-4, IL-6, IL-10, IL-17A, TNF-α, and IFN-γ levels was performed using the Cytokine Bead Array Th1/Th2/Th17 kit (BD Biosciences, San Jose, CA, USA). Samples were acquired with a FACS II LRS Fortessa (Becton, Dickinson and Company, USA) according to the manufacturer’s instructions and analyzed using the FCAP Array Software 3.0 (BD Biosciences, San Jose, CA, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!