EVs isolated from non-labeled hBM-MSCs were tagged with PKH26 (Red Fluorescent Cell Linker Kits MINI26; Sigma-Aldrich Co., St Louis, MO, USA) at room temperature (RT) for 5 min in the dark and blocked with fetal bovine serum (FBS), according to manufacturer’s instructions. The unincorporated labels were removed by EV centrifugation at 100,000×g for 75 min at 4 °C using a Thermo Scientific T-865 Fixed Angle Rotor Thermo Scientific Sorvall WX Ultracentrifuge Series. EVs were washed with DPBS and subjected to additional centrifugations. Then, the pellet was re-suspended in 1 ml DPBS for further use.
T 865 fixed angle rotor
Manufactured by Thermo Fisher Scientific
The T-865 Fixed Angle Rotor is a laboratory centrifuge component designed for use with compatible Thermo Fisher Scientific centrifuge models. The rotor features a fixed-angle configuration and is constructed for reliable performance during high-speed centrifugation procedures.
Automatically generated - may contain errors
Lab products found in correlation
Sorvall wx ultracentrifuge series, by Thermo Fisher Scientific (1 mentions)
Red fluorescent cell linker kits mini26, by Merck Group (1 mentions)
Wx ultracentrifuge series, by Thermo Fisher Scientific (1 mentions)
Sorvall wx ultracentrifuge, by Thermo Fisher Scientific (1 mentions)
Centrifuge 5804 r, by Eppendorf (1 mentions)
3 protocols using t 865 fixed angle rotor
1
Labeling of Extracellular Vesicles from hBM-MSCs
2
Fluorescent Labeling of Extracellular Vesicles
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EVs isolated from nonlabeled hBM-MSCs were tagged with PKH26 at RT for 5 minutes in the dark and blocked with FBS, according to manufacturer’s instructions. The unincorporated labels were removed by hBM-MSC-EVs centrifugation at 100,000× g for 75 minutes at 4°C using a Thermo Scientific T-865 Fixed Angle Rotor, Thermo Scientific Sorvall WX Ultracentrifuge Series. hBM-MSC-EVs were washed with DPBS and subjected to additional centrifugations. Then, the pellet was re-suspended in 1 mL of DPBS for further use.
3
Ultracentrifugation for CNT Dispersion
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Visual inspection of CNT dispersions after performing regular centrifugation (Eppendorf Centrifuge 5804 R, 2200 g, 30 min) showed that this was not sufficient to sediment the suite of CNTs tested. Therefore, ultracentrifugation was employed for further testing. Dispersions of each CNT type were placed in 36 mL centrifugation tubes and centrifuged at 40,000g for 30 min and 50,000g for 30 min; (T-865 Fixed Angle Rotor, Sorvall TM WX Ultracentrifuge, Thermo Scientific). After centrifugation, the supernatant was carefully removed using a glass pipette, taking care not to re-suspend settled CNTs. UV-vis absorbance measurements of the supernatant (800 nm) were the performed to assess the separation efficiency of CNTs from water phase.
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