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Immunoprecipitation of media samples was done using Protein-G agarose (Invitrogen).
The amount of glucose present in the media samples was determined using the Glucose Assay Kit (GAGO-20) from Sigma.
The mAb concentration in the different media samples was quantified by ELISA (Montgomery, TX, US) and UV vis spectroscopy using a Nanodrop Lite system (Thermo, Wilmington, DE, USA).
The amount of lactate present in the media samples was determined by generally following the Sigma Diagnostics Procedure No. 826-UV.
The IgG concentration in the different media samples was quantified by ELISA (Montgomery, TX, US) and UV vis spectroscopy using a Nanodrop Lite system (Thermo, Wilmington, DE, USA) prior to storage at −80 °C until further use.
The secretion of albumin and urea into media samples was determined 1 day after culture initiation and every 2 days thereafter using a human albumin enzyme-linked immunosorbent assay (ELISA) kit (Bethyl Laboratories) and QuantiChrom urea assay kit (Bethyl Laboratories).
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Media samples were collected from the cell culture dish, and 600 μl of the media sample was used for NMR analysis.
But he added that other tests of media samples were also being conducted at the same time.
Media samples were collected at 1, 2, 4, 8, 24, and 48 hours.
These results suggest that voids in post-treatment media samples were filled or occupied with nutrients and particulates adsorbed onto the media surface or biofilms, thus leading to the reduction of BET specific surface area and pore volume.
To test the validity and applicability of the previously established guidelines with regard to sample size, resolution and accuracy, the micro-structural details of representative porous media samples are acquired using x-ray micro-tomography.
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