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The mixture was vortexed for 2 min at maximal settings and centrifuged for 15 min at 17 000 g at 4 °C.
In order to facilitate RES inclusion in cyclodextrin, RES powder was added to the HPCD solution, vortexed for 5 min at maximal settings, sonicated for 1 h in an ice bath, and finally vortexed at 1200 rpm for 24 h at room temperature.
Nevertheless, repetitive mossy fiber stimulation (10 Hz for 1 s, 100 Hz for 1 s, or 10 Hz for 5 min) at maximal intensity did not affect GABA(A) receptor-mediated currents evoked by photorelease of GABA onto the proximal portion of the dendrite, where recurrent mossy fiber synapses were located.
The supernatant was trasferred to a new tube and centrifuged for 30 min at maximal speed in an Eppendorf centrifuge at +4°C.
Plasma was centrifuged for 5 min at maximal speed in a tabletop microcentrifuge.
For total protein analysis, samples were centrifuged for 15 min at maximal speed and supernatant was collected.
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Slides were heated for periods of 2 min at a maximal power setting (about 800 W) and for 2 × 2 min at the submaximal power setting (600 W).
DNA solutions were transferred to a 0.22 μm sterile Ultrafree-MC spin filter (Millipore) and centrifuged for 2 min at a maximal speed of 8000 × g to ensure the complete removal of live forms of B. anthracis from DNA.
Before testing, each subject performed an active warm-up with cycloergometer for 10 min at sub-maximal level.
Receptor-independent Ab uptake was very low (as seen in cells expressing a nontagged h.XGnRHR), but there was a clear agonist-dependent internalization of h.XGnRHR and mGnRHR that was measurable at 30 min and maximal at 60 min (Fig. 6).
On Study day 2, subjects had a muscle biopsy and blood sampling performed before and after KE exercise (60 min at 50% of maximal KE determine on day 1).
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