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The DEAE-cellulose column, previously conditioned with 0.2 M aq.
The product was obtained by filtration and then purified through a cationic exchange resin Dowex 50W X8–100 (50–100 mesh), previously conditioned with a 10% HCl solution.
Soxtec extracts were evaporated, ca. 1 mL, and additionally purified with a multilayered column previously conditioned with n-hexane (50 mL).
This solution was filtered and purified through a cationic exchange resin Dowex 50W X8–100 (50–100 mesh), previously conditioned with a 10% HCl solution.
For cation-exchange chromatography, Fr-LH-2 3 containing active components were applied to a 600 mL-bed volume oFr-LH-2 3llulose Fr-LH-2 3xcontainingrier, Wactivepen componentseviously conditioned were 0.2 M applied
Drug priming injections have been shown to reinstate CPP in animals previously conditioned with cocaine [18], morphine [19], amphetamine [20], nicotine [21] and ethanol [22].
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Briefly, hESCs were plated as single cells at a density of 18,000 cells/cm² in medium previously conditioned for 24 h with mitomycin C-inactivated mouse embryonic fibroblasts, containing 10 μM ROCK inhibitor Y-27632 and 10 ng/ml bFGF.
The target compounds were extracted in one step, by a method described elsewhere [ 18], using a Baker vacuum system (J.T. Baker, The Netherlands) and Waters (Milford, MA, USA) Oasis HLB cartridges (60 mg, 3 mL) previously conditioned at neutral pH with 5 mL methanol then 5 mL deionised water (HPLC grade).
To test this, pLB, pLUM or MCF-7 cells growing as three-dimensional colonies were incubated with media previously conditioned (CM) by pLB, pLUM or MCF-7 cells grown on plastic.
After 8 extinction trials, reinstatement of the CPP was observed following a nicotine priming injection (0.15 mg/kg, s.c). in adolescents that had previously been conditioned with 0.03 mg/kg nicotine; vehicle-treated rats did not show a significant preference for either compartment.
The different cartridges were previously activated and conditioned with 3 mL MeOH followed by 3 mL H2O.
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