Flow cytometry analysis was determined by analytical DNA flow cytometry. In this work, HepG2 and HT-29 cells were harvested and adjusted to 104 cells/ml (SPL, Korea) and then incubated for 6 hr with 10 µg/ml of peptide samples. The cells were centrifuged at high speed (12,000 rpm) for 20 sec. The pellet was washed with saline buffer, after repeating centrifugation, resuspended in 0.2 ml of lysis buffer (0.1% sodium citrate and 0.1% Triton X-100) containing 50 ?g/ml propidium iodide (PI), a highly water-soluble fluorescent compound, and stained with this reagent at 37 °C for 15 min in the dark. The cells were then evaluated for the DNA fragmentation analysis using a FACScalibur flow cytometry equipment (Becton Dickinson, CA, USA) supplied with the flowing software 2.5.1 (23 (link)).
Facscalibur flow cytometry equipment
Manufactured by BD
Sourced in United States
The FACScalibur is a flow cytometry instrument manufactured by BD. It is used for the analysis and quantification of cells, particles, and molecules in a fluid sample. The FACScalibur can measure multiple parameters of individual cells, such as size, granularity, and the expression of specific proteins or markers on the cell surface. This data can be used to identify and characterize different cell types within a heterogeneous sample.
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Lab products found in correlation
2 protocols using facscalibur flow cytometry equipment
1
Flow Cytometric DNA Fragmentation Assay
2
Cytokine Secretion Measurement in PBMCs
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From the supernatants of human PBMC cultures, cytokine secretion (IFN-γ, TNF, IL-2, IL-4, IL-6, IL-10) was measured using the BD™ CBA Human Th1/Th2 Cytokine kit (Becton Dickinson, United States). The reading was performed in the FACSCalibur flow cytometry equipment (Becton Dickinson, United States), according to the manufacturer’s guidelines. The results were analyzed using the FCAP Array 3.0 software (Becton Dickinson, United States), being normalized with the results obtained by non-stimulated cells. So, if the value is presented as negative, it means that there is an inhibition of cytokine production since the detected value is lower than that presented by the non-stimulated PBMCs.
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