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The thinnest part of the tibial component is maintained at 3.5 mm to keep some of the patient's own bone.
Depending whether open or closed KATP channels were required, 3 μM (for ATP-free pipette solution; this small amount of ATP was added to prevent the channel run-down) or 3 mM (to keep KATP channels closed of ATP) of ATP was added.
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FeSO4 solutions used in tetrahydropterin and phenylalanine binding experiments were prepared in 10 mM HCl to keep iron in its reduced state.
After a strip with adherent cells was clamped in the device, cells were covered with growth media plus 20 mM HEPES buffer (to keep pH stable outside the incubator) and covered with a 9×18 mm coverslip.
After drying in a speed vacuum centrifuge for 10 minutes, the gel slices were rehydrated for 15 minutes at 4°C in 25 ul trypsin (Sigma, Proteomics Grade) in 25 mM ammonium bicarbonate (pH 8-8.5) and overlaid with minimum volume of 25 mM ammonium bicarbonate to keep them immersed throughout the digestion process and incubated for 16-24 h at 37°C.
Furthermore, the gap sizes should be smaller than 3 mm in order to keep losses small.
The nozzle with the 60° opening angle and 0.8 mm bore allowed to keep molten silicates in a stable position for up to >1 h (Fig. 4).
All three specimens were designed with the same minimal thickness of the contralateral cortical of exactly 5 mm in order to keep the highly stressed material volume identical or at least as comparable as possible.
Membranes were prepared from HEK293 cells transfected with the receptors using PBS and 5 mM EDTA, which helps to keep free cysteine residues in the reduced state.
80 μl of cell pellets were mixed with 70 μl 100 mM Triethylammoniumbicarbonate (TEAB) solution to keep the pH stable at about 8. 50 μl glassbeads was added and the sample was mixed vigorously by Fast Prep (MP Biomedicals Solon, OH, USA) to break the cells (20 seconds at speed 6 for 4 times, kept 30 seconds on ice between every run).
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