(F2442, Sigma-Aldrich) in dPBS before FACS purification. This was done for samples collected from four different fish for instrument operation, data acquisition, and analysis. A 637 nm and a 488 nm laser
Following 30–60 min of incubation, the cells were gently washed with RPMI 1640, passed through another 70 μm cell strainer into a new 50 ml centrifuge tube, and pelleted by centrifugation at 1500 rpm, RT for 5 min. The cell pellet was resuspended in 2–3 ml PBS + ...
7. Wash the cells once: fill the flow tube with staining buffer, centrifuge the sample to pel- let the cells, and then decant the supernatant. Intracellular staining 8. Permeabilize the cell sample by adding 100 µl FIX & PERM® Reagent B. 9. Prepare Zenon® complex with primary ...
To pellet PEVs, the supernatant was carefully transferred to another tube and centrifuged 22,000× g for 45 min [58]. Subsequently, the pellet was aliquoted in dPBS and frozen at −80 °C until use. All centrifuge steps were made at 4 °C. Extracellular vesicle protein concentration was...
choices.Thebeamgeometryisoptimizedfor mosthematopoieticstemcells.Thetophat designofthe375-nmNearUVlasertriples theamountoflightatthesampleintercept. The375-nmNearUVlasercanbeoperated togetherwiththeredandbluelasers. UpgradesfromBDFACSAria toBDFACSAriaII ...