The size distribution and particle concentration of fractions recovered after ODG were measured using tunable resistive pulse sensing (TRPS) by qNano (Izon, Christchurch, New Zealand) following the manufacturer’s instructions for working with smaller range nanopores. Voltage and pressure values were set to optimize the signal to ensure high sensitivity. A nanopore NP100 was used for all fractions analyzed except for the grape vesicles, where a NP150 was used. Calibration was performed using CP100 carboxylated polystyrene calibration particles (Izon) at a 1:1,000 dilution. Samples were diluted 1:5 and applied to the nanopore. The size and concentration of particles were determined using the software provided by Izon (version 3.2). Protein concentration was measured in all fractions, and EV purity determined as described previously (35 (link)).
Cp100 carboxylated polystyrene calibration particles
Manufactured by Izon Science
The CP100 carboxylated polystyrene calibration particles are a laboratory product designed for calibration and standardization purposes. These particles are spherical in shape and have a carboxylated surface, which provides a consistent and uniform surface chemistry. The particles are available in a range of sizes, allowing for precise calibration and measurement across various applications.
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Lab products found in correlation
4 protocols using cp100 carboxylated polystyrene calibration particles
1
Nanoparticle Characterization by TRPS
2
Measuring Extracellular Vesicles by TRPS
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Tunable resistive pulse sensing (TRPS) was employed using a qNano system (Izon Science) to measure the particle concentration and size distribution of Evs. Briefly, a nanopore NP150 or NP400 (for 12k and 15k Evs, respectively) was used, and pressure and voltage values were set to optimize the signal to ensure high sensitivity. All samples were diluted (1:10 for OvEVs and 1:20 for grapesomes) before applying to the nanopore, and CP100 carboxylated polystyrene calibration particles (Izon Science) were used for calibration. Size distribution and concentration of particles were analyzed using the software provided by Izon (version 3.2).
3
Particle Size and Concentration Analysis via TRPS
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The size distribution and particle concentration of fractions recovered after ODG were measured using tunable resistive pulse sensing (TRPS) by qNano (Izon, Christchurch, New Zealand) following the manufacturer’s instructions. Voltage and pressure values were set to optimize the signal to ensure high sensitivity. A nanopore NP100 was used for all fractions analysed except for fraction 9, where a NP150 was used. Calibration was performed using CP100 carboxylated polystyrene calibration particles (Izon) at a 1:1000 dilution. Samples were diluted 1:5 and applied to the nanopore. The size and concentration of particles were determined using the software provided by Izon (version 3.2).
4
Vesicle Protein Content and Size Analysis
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Vesicle protein content was determined by BCA (Bio-Rad). The particle concentration and size distribution of each fraction collected after (i) ultracentrifugation only and (ii) iodixanol gradient was determined by TRPS using a qNano instrument (Izon Science, New Zealand) following the manufacturer’s instructions. Briefly, a nanopore NP150 was used, and pressure and voltage values were set to optimize the signal to ensure high sensitivity. Samples were diluted 1:5 in electrolyte (Izon Science) before applying to the nanopore. Calibration was performed using CP100 carboxylated polystyrene calibration particles (Izon Science) at a 1:2,000 dilution. Size distribution and concentration of particles were analyzed using the software provided by Izon (version 3.2), and purity analysis (number of particles/µg of protein) was performed following established methods (Webber and Clayton, 2013 (link)).
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