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In the ESI source, capillary voltage, nebulizer pressure, dry heater, and flow rate of dry gas were set to 4500 V, 2.0 bar, 350 °C, 220 °C and 10 L/min, respectively.
In the APCI source, nebulizer pressure, capillary voltage, dry heater, and flow rate of dry gas were set to 3.0 bar, 3000 V, 250 °C and 2.5 L/min, respectively.
The samples were incubated at 97°C for 30 min on a dry heater block, cooled for 5 min and centrifuged 10 000 × g for 10 min at room temperature.
The extraction was quantified, diluted in 1 × NuPAGE/LDS loading buffer (Invitrogen), heated in 95 °C dry heater (Labnet, Woodbridge, NJ, USA) for 5 min, and loaded in 10% NuPAGE/Bis/Tris gel (Invitrogen) with NuPAGE/MES/SDS buffer (Invitrogen).
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Extraction was carried out at 80°C in a dry block heater for 1 h.
The optimized MS operating conditions were as follows: capillary voltage 4500 V, nebulizer gas pressure 0.8 bar, drying gas flow rate 8 l/min, dry gas heater temperature 200 °C in the positive ion mode (ESI+).
A 300 μL of dansyl chloride solution was then added, and solution was mixed vigorously; then, tubes were kept in dry block heater at 40°C for 30 min. The tubes were then cooled, and the dansyl derivative was extracted three times with 1.5 mL of dichloromethane by a vortex mean.
Samples were then heated at 95°C in a dry block heater for 5 min, cooled and loaded into 12% Bis-Tris NuPAGE gels.
For the incubation of the glass vials, a dry-block heater Type BS25 230D (Aerne Analytic, Pfaffenhofen, Germany) was used.
Figure 2 shows the essential features of the dryer consisting of the solar collector (air heater), drying cabinets, and drying trays.
Do not use dry heat such as heaters, fireplaces or heating pads.
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