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Immunolabelling was visualised and densitometry performed using an Odyssey system (LiCor, Cambridge, UK).
The antigen-antibody complex was visualized and quantitated using an Odyssey system (LI-COR, Lincoln, NE, USA).
For loading control the blot was incubated with a monoclonal anti-mouse beta-actin (Sigma Aldrich) and an anti-mouse Alexa Fluor 680 secondary antibody (Life Technologies), then imaged using an Odyssey system (LI-COR Biosciences Ltd., Cambridge, Cambs, UK).
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Bands were visualized using an Odyssey imaging system (Li-COR biosciences, Lincoln, NE).
Blots were analyzed using an Odyssey imaging system.
Immunoreactive protein bands were detected using an Odyssey Scanning System (LI-COR Inc., Superior St., Lincoln, NE, USA).
In some experiments, the 'active' conformation of the G protein was stabilized by replacing GDP in the binding and wash buffers with 30 µM GTPγS or a mixture of 30 µM GDP/30 µM AlCl3/10 mM NaF. Immunoblot quantification was performed by infrared imaging following the manufacturer's protocols using an Odyssey imaging system (Li-Cor Biosciences).
For analysis of these levels and those of LC3-II levels in PC12 cells treated with rilmenidine, western blots were probed with secondary antibodies conjugated to IRDye® for detection at 780 or 680 nm (Li-Cor Biosciences) and visualized and quantified using an Odyssey imaging system (Li-Cor Biosciences).
The images were acquired using an Odyssey infrared imaging system (LI-COR, USA).
Signals were imaged using an Odyssey infrared imaging system (Licor Biosciences).
The precipitates were analyzed by Western blot with the indicated antibodies and visualized by incubation with IRDye800-conjugated secondary antibodies (diluted 1∶10,000) using an Odyssey infrared imaging system (LICOR Inc., Germany).
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