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The 4-helix bundle structure present in aqueous detergent solution and at the air-water interface, is preserved in multilayer films of hbAP-PheCN, enabling vibrational spectroscopy of both the protein and its nitrile label (-CN).
There, samples were removed from the plastic bags, washed well with a de-ionized aqueous detergent solution (Triton X-100) and dried before cutting and weighing.
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Ultrasonic technology is commonly used to provide mechanical force to augment the aqueous detergents.
Decellularization by extended exposure to aqueous detergents can damage the microstructure or deposit cytotoxic residue.
Equal volumes of aqueous and detergent phases were analyzed by SDS-PAGE and western blot.
Aliquots of the aqueous and detergent extracted protein fractions were flash frozen in liquid nitrogen and stored at -80°C.
The aqueous and detergent phases from this procedure were adjusted to the equal volume with 10 mM Tris/HCl (pH 7.4) and 150 mM NaCl plus protease inhibitors.
These aggregates may consist of oligomeric complexes of non-native secondary structures and demonstrate poor solubility in aqueous or detergent solvents [ 3].
The approximate relative amounts of CD59 in the aqueous and detergent phases were found to be 20 and 80%, respectively, as judged from the intensities of staining (n=7).
Although the amount of protein solubilized in the aqueous or detergent buffers were similar between young and old worms, FA treatment of the SDS-insoluble fraction yielded significantly more proteins from old worms when compared with young adults (Fig. 1).
After 3 months of treatment, the cortex was sequentially extracted with RAB (aqueous buffer), RIPA (detergent buffer), and finally 70% FA to solubilize the detergent insoluble final pellet.
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