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Each separation step was monitored by TLC on silica gel plates (GF254, Qingdao Marine Chemical Co., Ltd., Qingdao, China), and spots were visualized using Dragendorff's reagent.
UV spots were visualized using ultraviolet irradiation (at 254 365 nm) and by spraying with 10%H2SO44, followed by heating with a heat gun.
For TLC analysis, a silica gel (Merck, Kieselgel 60) was used and spots were visualized using UV (254 and 360 nm) and sulphuric acid solution (10%) with after heating.
The limits of detection of separated amino acids were determined by spotting 0.10 μL of amino acids solutions on the TLC plates which were developed with mobile phase M11 and the spots were visualized using ninhydrin.
Spots were visualized using an alkaline phosphatase conjugate substrate kit (Biorad, Hercules, CA, USA) and enumerated using an automated ELISpot reader.
Reactive spots were visualized using the Western Lightning Chemiluminescence kit (Perkin-Elmer Life and Analytical Sciences, Woodbridge, Ontario) and images captured/transferred onto BioMax Film Perkin-Elmer Life and Analytical Sciencess, Woodbridge, Ontario).
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Growth of these bacterial pathogens in the inoculated spots was visualized using UV light under dark.
The spots of individual separated compounds were visualized using UV light, and their identities were confirmed with a matrix-assisted laser desorption/ionization time of flight mass spectrometry.
Spots corresponding to positive peptides were visualized using the ECL system.
Positive spots corresponding to PP2A-binding peptides were visualized using the ECL system.
After washing away cells, spots corresponding to B cells secreting antibody were visualized using HRP conjugated anti-mouse Ig and AEC substrate, and counted using a stereo dissection microscope.
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