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Specific activity, activity recovery, purification fold, volume ratio, enzyme and protein partition coefficients were considered as response variables.
The flow-through and fractions from washing and elution were collected and precipitated with 4 fold volume of acetone for 2 h at −20°C.
Following centrifugation at 100,000 g (4 °C; 30 min), supernatants collected and neutralised with a19 fold volume of neutralisation buffer (1 M Tris base, 0.5 M Na2HPO4, 0.05% NaN3).
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Proteins were then eluted with 5-fold volume of glutathione solution (Sigma-Aldrich), and were condensed using 10 kDa protein concentrators (Thermo Fisher).
We found that cereal straw could be completely dissolved in a 10-fold volume excess of trifluoroacetic acid (≥99%) (TFA).
To date, however, only a five-fold volume expansion is achievable in a single-use stirred tank bioreactor, which in turn, necessitates intermediate equipment to scale up the culture to production volume.
DNA was precipitated by addition of the 0.7-fold volume of isopropanol.
DNA was precipitated by addition of the 0.7-fold volume of isopropanol to the aqueous phase.
DNA was precipitated by addition of a 0.6-fold volume of isopropyl alcohol followed by washing in 70% ethanol to remove basic ions.
The amide was repetitively precipitated with 10-fold volume of diethyl ether and washed with THF till no ethanolamine in ether solution was detectable by reaction with TNBS.
The mixture was diluted to two-fold volume with methanol in case the color was too dark, the peroxide value was measured by monitoring absorbance at 500 nm in a spectrophotometer (Unico 2100, Shanghai, China).
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