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Here, we employed SRM assay for verification of SILAC data, since we have previously validated its accuracy and effectiveness for verification of candidates in amniotic fluid [ 15].
To facilitate development of a LC-SRM assay for verification of the exploratory iTRAQ findings, we first compiled an MS/MS spectra library of dimethylated peptides from which appropriate surrogate peptides were selected.
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In detail, 96 well ELISA plates (Dynex Corporation, Chantilly, VA, USA) coated with the various proteins were used in biochemical assays for verification of protein:protein interaction.
Two candidate protein biomarkers were arbitrarily chosen based on fold differences (Table 1), putative roles in the pathogenesis of prostatitis, and the availability of assay reagents for verification using the gold standard in protein quantification, the enzyme-linked immunosorbent assay (ELISA).
Antigen coating was performed as described above (Enzyme linked immunosorbent assay (ELISA) for verification of immunogenic proteins).
For the scFv titration ELISA the antigen was coated as described above (Enzyme linked immunosorbent assay (ELISA) for verification of immunogenic proteins).
A paper describing the conceptual idea of using passive neutron assay for the verification of large size uranium samples in fuel fabrication plants was first presented at the 2001 ESARDA conference.
This example indicates one of the drawbacks for application immune-based assays for protein verification: lack of specific antibodies.
We also designed targeted SRM assays for candidate verification and identified two proteins (SOD1 and NES) that could be involved in the molecular pathogenesis of DS during fetal development.
TaqMan Copy-Number assays were used for verification of CNVs showing significant association with HNPCC/LS identified by both software programs.
For verification assays, transduced hMSCs were seeded in 96-well plates.
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