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In the present study, several analogs of BTB 06237 were synthesized and analyzed for activity against Leishmania axenic amastigotes, their ability to reduce the level of parasitemia in peritoneal macrophages, and their ability to generate reactive oxygen species (ROS) in L. donovani promastigotes.
To evaluate thermostability, three replicates of sonicated suspensions were maintained at RT, 50°C, 60°C or 70°C for 20 min before being transferred to ice and analyzed for activity.
Aliquots were removed at specified time intervals and analyzed for activity on 12% native-PAGE and stained for SOD activity.
Sera from 22 RRMS patients and 66 controls from the USA's National Institute of Health (NIH) were analyzed for activity to HHV-6 [ 24].
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Serum samples were analyzed for activities of antioxidant enzymes including GSH-Px and for TBARS using the commercial kits provided by Nanjing Jiancheng Bioengineering Institute (Nanjing, China) [ 20].
Liver homogenates were analyzed for activities of the following enzymes: glutathione peroxidase (GPX), glutathione-S-transferase (GST), and NAD(P H: quinone reductase (QR).
For activity-based screening, super pools were analyzed for chitinolytic activity using the activity assay with 4-methylumbelliferyl-N-N′-chitobiose as substrate, as described below.
The fractions with the higher ASNase II activity were pooled and analyzed for total activity (U), total protein level (mg), and specific activity (U/mg).
The reconstituted HqD complex(s) were subsequently analyzed for HqD activity using a spectrophotometric activity assay.
Protein fractions were dialyzed into unlinkase buffer and analyzed for unlinkase activity; fractions containing peak activity were then used in further assays.
Activity specific bias was analyzed for each activity separately (the 15 time intervals coded #1- 15).
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