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Furthermore, we've identified DAPI as a reliable and very sensitive stain that allows for the detection of the pyrophosphate moiety.
The following day the medium was removed from the wells, the cells fixed with 0.5% glutaraldehyde (Sigma-Aldrich), washed twice with 2 mM magnesium chloride (Sigma-Aldrich) in PBS and stained with an X-gal sensitive stain (10 mM potassium ferricyanide/ferrocyanide, 0.02% Tergitol, 0.01% sodium deoxycholate, 0.5 mg/ml X-gal, 2 mM magnesium chloride in PBS).
However, as supported by several studies [ 10, 11] calcofluor white is indeed a highly reliable and sensitive stain for fungal detection under fluorescence microscope.
Therefore, using sensitive stain increases protein sample dynamic range, leads to successful gel imaging, and finally leads to successful mass spectrometric identification and immunological validation [ 11].
Confocal microscopy images of cells stained with Nile Red – a lipophilic dye that serves as a sensitive stain for the detection of neutral lipids (Greenspan et al, 1985) – demonstrated the presence of lipid droplets in the cell cytoplasm.
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Immunohistochemical staining was accomplished by rocking slides using the Antibody Amplifier™ to ensure even, consistent and sensitive staining.
In this work we used a more sensitive staining approach than the one we used previously [ 7] which allowed a better characterization of both the proteome and phosphoproteome of a susceptible cultivar.
While a number of sensitive staining methods have been developed for protein quantification in solution as well as on solid supports [ 27], these principles have not been extended and generalized for fluorescence microscopy.
Given the high sensitivity of the NAF-1 antibody, we were able to perform a sub-cellular localization study of Kir6.1 protein in the mouse heart (Fig 4; this approach was not possible with other antibodies, which produced less sensitive staining).
After time-sensitive stain development, sections were counterstained in hematoxylin (Sigma) for identification of cell nuclei.
We next examined whether the UBB+1-induced loss of clonogenicity correlated with the manifestation of oxidative stress, which can be detected by the intracellular conversion of the reactive oxygen species (ROS -sensitive stain dihydROS -sensitiveE) to fluorestain ethidihydroethidium
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