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Methotrexate and 6-mercaptopurine (both Sigma) were dissolved in NaOH at a maximal final concentration of 50 mM (Sigma).
Maximal final concentration of CM used in the experiments was 75%%.
AR-R 17779 was dissolved in Dimethyl Sulfoxide (DMSO) and then diluted in culture media to a maximal final concentration of 3.3% DMSO.
Epirubicin was reconstituted with 0.9% normal saline or sterile water for injection to produce a maximal final concentration of 2 mg ml−1.
Rat BNP-(1 32) and C-ANF(4 23) were obtained from Sigma-Aldrich UK, and both CNP-(1 22) and 8Br-cGMP from Tocris bioscience UK. sKATP channel opener pinacidil (Sigma-Aldrich, UK) was dissolved in dimethylsulfoxide (DMSO; maximal final concentration of 0.25 % v/v).
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Stock solutions of the compounds were diluted in culture medium to a maximal final DMSO concentration of 0.3% for PHH spheroids and 0.6% for HepaRG spheroids.
The maximal effect was obtained using a final concentration of 10-20 nM (Fig. 6E), in agreement with values reported in the literature (Cheung and Wong, 2008).
To determine the maximal fluorescence of the cells, ionomycin (final concentration 2 μ m) was added between the 40th and 41st frames.
The addition of FCCP (final concentration 2 μM) gave maximal respiration, whereas 1 mM cyanide elicited minimal respiration.
The therapeutic drugs were added to achieve a final concentration of 3× the maximal plasma concentration (as defined by the CLSI EP7-A2 Guideline) [ 15], if known.
In one control well, NP40 was used at a final concentration of 0.1% to determine maximal LDH release.
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