Plasma samples were filtered (Minisart NML syringe filters, pore size: 0.8 μm; Sartorius) and cleaned from debris by two serial centrifugation steps: 1.500 × g for 10 min followed by 10.000 × g for 10 min before loading on a pre‐rinsed qEV2/70 nm column (Izon Science LTD, Cat. No. SP4). EV were collected from 1st to 5th fractions by using an Izon Automatic Fraction Collector (AFC) and concentrated in Amicon Ultra 15 filtering units (Merck Millipore, Cat. No. UFC910024) by centrifugation at 3.000 × g for 20 min on a 5810R benchtop centrifuge (Eppendorf). The volumes eluted between the 6th and the 20th fractions were collected and processed for cfDNA isolation as previously described (Lee et al., 2018 (link)).
5810r benchtop centrifuge
Manufactured by Eppendorf
The 5810R is a benchtop centrifuge designed for a wide range of laboratory applications. It features a high-performance refrigeration system and can accommodate rotors for various tube sizes. The centrifuge provides consistent performance and reliable operation.
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Lab products found in correlation
2 protocols using 5810r benchtop centrifuge
1
Extracellular Vesicle Isolation from Plasma
2
Hexapeptide NV Binding with Microtubule
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Example 6
Hexapeptide NV Binding with Microtubule at Tubulin Polymerization
Mixture 1: Alexa Fluor 568-labeled tubulin (0.5 μL, 15 mg mL−1, 65% labelling ratio), Tubulin (2 μL, 20 mg mL−1), FITC-NV (0.5 μL, 1 mM), GTP (0.2 μL, mM) and BRB80 (46.8 μL; 80 mM PIPES, 1 mM EGTA; 1 mM MgCl2, pH adjusted to 6.8 by using concentrated KOH solution) were mixed on ice. The mixture was incubated for 30 min on 37° C. water bath.
Mixture 2: Taxol (0.4 μL, 1 mM) and BRB80 (180 μL) were mixed.
Final Mixture: After 30 min incubation 50 μL warmed mixture 2 was added into mixture 1. Then this mixture was separated in a table top centrifuge (7 min, 12000 rpm, Eppendorf 5810R bench-top centrifuge, rotor type F-34-6-38). The coloured pellet was resuspended in warm mixture 2. 10 μL resuspended solution of microtubule was placed on glass cover slip and imaged under inverted fluorescence microscope (NIKON Ti-U) in 40× magnification using ANDOR iXON3 camera.
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