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Following washes and 30 min incubation with HRP-conjugated goat anti-rabbit or goat anti-mouse secondary antibodies (Jackson Labs), membranes were visualized by an enhanced ECL Renaissance kit (NENTm Life Science Products) and then exposed to Hyperfilm ECL (Amersham) for various times to get bands within the linear range of sensitivity.
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Blots were imaged and quantified by assessing peak density after ensuring bands were within the linear range of detection using the Chemidoc XRS system (Bio-Rad, Hemel Hempstead, UK).
Serial dilution of samples for immunoblotting confirmed that the density of bands was within the linear range of detection.
The relative expression levels were normalized using tubulin bands within the same linear range of detection.
The relative expression levels for the dystrophin bands were normalized using tubulin bands within the same linear range of detection.
PCR analysis of ChIP samples used 1 50 and 1 250 dilutions of the ChIP DNA to obtain band intensities within the linear range of the input DNA dilutions.
The intensity of the developed bands was well within the linear range of detection.
Appropriate exposures of the film were used to ensure that band intensities were within the linear range.
The final reaction products were developed using SuperSignal West Pico Chemiluminescent Substrate for 5 min and the band intensities were within the linear range of detection.
The net pixel value for each protein band that lied within the linear range of detection was normalized to the co-analyzed standard.
Dispersion effects forbid wave transmission within the stop band of the linear periodic structure.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com