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Prior to B-phycoerythrin purification, the protein adsorption of the vortex flow reactor was characterized through hydrodynamic studies and a dynamic capacity measurement using a standard protein.
Protein concentrations were determined using a standard protein assay (Bio-Rad, Hercules, CA) with bovine serum albumin as the standard.
Alternatively, cells were harvested in NP-40 lysis buffer (100 mM Tris (pH 8.0), 100 mM NaCl2, 1% NP-40) containing an EDTA-free protease inhibitor cocktail (Boehringer Ltdnheim-Roche Diagnostics Ltd, Burgess Hill, UK) to determine total protein concentration using a standard protein assay (Biorad, Hemel Hempstead, UK).
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Using a standard protein-ligand docking program with only minor modifications, four new ligands with binding affinities in the micromolar range were identified, including two compounds based on molecular scaffolds not resembling known ligands.
The concentration of the recombinant protein was determined using a standard BCA protein assay kit (Pierce).
Total protein levels were measured using a standard Lowry protein assay.
Protein concentrations were determined using a standard Bradford protein assay, and preparations were quick-frozen in liquid nitrogen for short-term storage.
In some instances, mediator levels were normalized to protein content which was determined using a standard Bradford protein assay.
The predicted G. pallida protein set was first analysed using a standard secretory protein identification protocol.
The activity was normalised to the protein content, which was determined using a standard colourimetric protein assay (Sigma, Dorset, UK).
The protein concentration of secretomes was determined using a standard Bradford protein assay (BioRad, Hercules, CA, USA).
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