Blood samples (250 µL of whole blood) were collected in tubes with 3% potassium EDTA. After lysis of the red blood cells (twice) using ACK buffer (0.15 M NH4Cl, 10 mM KHCO3, and 1 mM EDTA monosodium; pH: 7.3) for 15 and 5 min separately, followed by washing with PBS (twice), the cells were simultaneously stained for effector T cells. Cells were surface-stained using the following anti-rat antibodies: anti-CD45-FITC (OX-1, Thermo Fisher Scientific, Waltham, MA, USA), anti-CD4-BV-786 (OX-35, BD Biosciences, San Jose, CA, USA), anti-CD8α-PerCP-e710 (OX-8, Thermo Fisher Scientific), and anti-CD62L-PE (OX-85, SONY) for CD4/CD8 T cells panel. The cell suspensions were analyzed by flow cytometry (BD LSR II including an FACSFlow Supply of the High Throughput Sampler System, BD Biosciences).
Facsflow supply of the high throughput sampler system
Manufactured by BD
Sourced in United States
The FACSFlow Supply is a core component of the High Throughput Sampler System from BD. It provides a consistent flow of sheath fluid to the flow cytometer, which is essential for the proper operation and performance of the system.
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Lab products found in correlation
2 protocols using facsflow supply of the high throughput sampler system
1
Effector T Cell Immunophenotyping
2
Cell Cycle Analysis by Flow Cytometry
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Identification of cell division stages is based on the different DNA amounts in these stages. DNA is stained with propidium iodide. A possible RNA interference is excluded by the RNase treatment. Cells were harvested, washed by phosphate buffered saline (PBS) and divided into equal aliquots (2•10 6 cells per sample) which were centrifuged, PBS was removed and cells were fixed by 1 ml of 70 % ethanol and left in the refrigerator for 2 h. Then, fixed samples were again washed by PBS, centrifuged at 850 × g and supernatant was removed. Fixed cells were finally treated with 50 μg•ml -1 RNase A for 15 min and then stained by 50 μg•ml -1 propidium iodide. These suspensions were measured on a 16 color flow cytometer BD LSR II including a FACS Flow Supply of the High Throughput Sampler System (BD Biosciences, San Jose, CA, USA) at excitation at 535 nm and emission at 617 nm. The percentage of each cell cycle phases was calculated using the single cell analysis software FlowJo.
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