The Cytometric Bead Array technique was employed to measure eight cytokines [38 (link)] (IL-1β, IL-6, IL-8, IL-10, IL-12p70, IFNγ, RANTES and TNF) in the collected supernatant at the study time points of 10 minutes, 2hours and 18hours, to determine how E2 concentrations affected the responses of the cultured HECECs to stimulation with TLR2 or TLR4 ligands. Each set of experiments consisted of five samples and the baseline cytokine expression levels were determined from the supernatant fluid of HECECs stimulated with just the relevant TLR ligand. Additionally, cytokine expression profiles of non-treated HECECs were used as control. Master Buffer Kit (558264, The BD™ CBA Human Soluble Protein Flex Set System, BD Biosciences) was used for these assays and the samples were run on the BD FACS Array flow cytometry machine, using FACP Array software.
Facs array flow cytometry machine
Manufactured by BD
The BD FACS Array is a flow cytometry machine designed for cell analysis. It utilizes laser technology to detect and analyze the physical and chemical characteristics of cells or particles in a fluid sample. The core function of the BD FACS Array is to provide researchers and clinicians with precise data on the properties and composition of complex cell populations.
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Lab products found in correlation
2 protocols using facs array flow cytometry machine
1
Cytokine Profiling in HECEC Cultures
2
Cytokine and Chemokine Profiling of Monocyte-HUVEC Interactions
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The cytometric bead array (CBA) technique was used to measure the inflammatory cytokines (IL-1 and IL-6), the anti-inflammatory cytokine (IL-10), and chemokines (IL-8 and MCP-1) within conditioned medium collected from monocytes and the HUVECs cultures. This assay was performed as previously described using kits from BD Biosciences, a BD FACS Array flow cytometry machine and FCAP Array software. The IL-1 and IL-10 cytokines were measured in the undiluted aliquot (30μl). However, IL-6, IL-8 and MCP-1 cytokines were above standard levels for undiluted aliquots. Consequently, serial dilutions (1:10 up to 1:200) were attempted to optimize the desired dilution. Thereafter, according to the optimum concentration within the sample, either 1:50 or 1:100 dilutions were applied to the samples, and each sample was multiplied according to its dilution factor before analysis was initiated.
Additionally, The CBA technique was used to measure the VEGF and sVCAM-1 in the supernatant medium collected from monocytes-HUVECs mono-or co-cultures.
Additionally, The CBA technique was used to measure the VEGF and sVCAM-1 in the supernatant medium collected from monocytes-HUVECs mono-or co-cultures.
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