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This assay was then scaled up to a 96-well plate format for screening MarsCy1 DRY mutant expressing cells against six characterized D2R antagonists.
We have developed a new and fast method to produce scFv antibodies by using only 5 μg of cell-free expressed protein and a naive scFv antibody library in combination with an antibody microarray format for screening.
Initially, four commercially available cell-based assays were selected to evaluate the suitability of the qHTS approach in the 1,536-well 1,536-well screening a non-drug–like compound library.
During Tox21 Phase I (proof of principle), many biochemical and cell-based assays were successfully developed and miniaturized in a 1,536-well 1,536-wellat for screening against the initial Tox21 collection of aplateimately 3,000 compounds in a quantitative high throughput screening (qHTS) platformatShukla et al. 2010; Tice et al. 2013).
Due to its quicker and easier production, it provides a robust format for screening of large numbers of antibody candidates, and can be converted to full IgG afterwards.
To address this specific need, in this study we have developed a novel, sensitive and quantitative luminescence based assay that does not involve the use of radioactive isotope, uses relatively low number of Treg cells, and can be easily modified to a high-throughput format for screening purposes.
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We developed three different assay formats for screening compound libraries.
The general format was designed for screening the small molecular compound library and identifying novel drug resistant inhibitor.
DNA from individual M2 mutant lines arranged in indexed 96-well plate format was used for screening.
This assay has been suitably converted to a high throughput microtitre plate format that is suitable for screening potential inhibitors.
One major advantage of the FRET assay over the SDS-PAGE-based assay is that it can be easily configured to a high-throughput format for analysis or for screening.
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