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Each sub-fraction obtained was tested for vasorelaxing activities and the most active fraction was subjected to further studies.
Chloroform fraction was most active fraction as it showed considerable activity against all bacterial strains.
Each dialyzed fraction was analyzed using caseinolytic method [ 23] for its PI activity and 500 μl of most active fraction (30-60%) was subjected to size exclusion chromatography using Sephadex G-75 (Sigma chemicals) gel filtration column (20 × 300 mm).
During early FPLC-separation steps (e.g. first Mono-Q separation), the elution profiles showed up to four heme peaks with veratryl alcohol oxidizing activity; we focused our purification efforts always on the most active fraction (i.e. that with the highest specific activity).
A bioguided assay fractionation was performed in order to obtain the most active fraction that could be further analyzed using HPLC and LC-MS methods to identify the bioactive compounds present in the fraction.
The most active fraction in each separation step was further purified and an active compound was isolated.
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We are currently busy in isolating and characterizing the constituents responsible for the antimicrobial activity from the most active fractions.
These fractions were tested for their antimicrobial/antioxidant activities and the most active fractions were further subjected to purification in order to isolate the active principles.
Further antiviral activities of the most active fractions at the highest dose were tested at days 1, 3, and 5. Analysis of inhibition of HBsAg and HBeAg secretions in the culture supernatants was done as mentioned above.
Antifungal activity was checked and most active fractions were used for further analysis.
We identified those compounds of the most active fractions that displayed anti-proliferative activity.
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