For measurement of soluble protein aggregates, at least three grids per protein were mounted, and at least three images per protein were analyzed. Length, defined as the long axis of each fragment measured, was measured using FIJI [77 (link)] and binned into 8 nm segments [66 (link)] using the scale bar as a standard (2.58 pixels/nm). Particles were excluded from analysis if they were on the border of the image or did not have clearly defined edges (i.e. due to a clustering). All particles meeting these criteria within the image field of view were also analyzed for width, defined as the short axis of each measured fragment.
Qualitative filter paper
Qualitative filter paper is a type of laboratory filtration paper used to separate solid particles from liquids. It is designed to remove fine particulates from solutions, facilitating the collection and analysis of the filtered material.
3 protocols using qualitative filter paper
Quantification of Protein Aggregate Size
For measurement of soluble protein aggregates, at least three grids per protein were mounted, and at least three images per protein were analyzed. Length, defined as the long axis of each fragment measured, was measured using FIJI [77 (link)] and binned into 8 nm segments [66 (link)] using the scale bar as a standard (2.58 pixels/nm). Particles were excluded from analysis if they were on the border of the image or did not have clearly defined edges (i.e. due to a clustering). All particles meeting these criteria within the image field of view were also analyzed for width, defined as the short axis of each measured fragment.
Synthesis of Cellulose Lignin Particles
acetone/water 3:1 (w/w) mixture and stirred for 3 h at room temperature.
The lignin solution was then filtered using a glass fiber (GF/F Whatman,
pore size 0.7 μm) to discard the undissolved aggregates. Then
270 g of water was poured rapidly into the filtered solution. The
resulting CLPs dispersion was placed in dialysis tubes (Spectra/Por
dialysis membrane, pore size 6–8 kDa) and dialyzed against
water for 2 days to remove acetone and low-molecular-weight impurities.
Afterward, the dispersion was concentrated under reduced pressure
at 45 °C, followed by filtration using VWR qualitative filter
paper, 415 (particle retention 12–15 μm). The final concentration
of CLPs dispersion was 3.83 wt %. The particle size distribution and
zeta potential of CLPs dispersion was measured by a Zetasizer NanoZS90
instrument (Malvern Instruments Ltd. U.K). The average size of CLPs
was 244 ± 4 nm, and the average zeta potential was −37
± 3 mV.
Extraction and Fractionation of Medicinal Plants
A fraction of each raw extract was then suspended in methanol:water, 9:1 and extracted with n-hexane. The residue was then suspended in distilled water and extracted successively with dichloromethane and ethyl acetate (analytical grade, VWR International, Milan, Italy).
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