HEK293T cells were gift of Colin Johnson at Oregon State University. The cells were maintained in Dulbecco’s modified Eagle’s media (Corning) supplemented with 10% fetal bovine serum (VWR) and penicillin/streptomycin (Corning). Subculture of cells was done twice a week using 0.05% trypsin/0.053 mM EDTA (Corning). HEK293T cells were plated in a white 96-well plate (Greiner) at a density of 3,000 cells per well and incubated for 48 h with 3-nitrotyrosine, or vehicle at concentrations indicated in the figure. Cell viability was measured using CellTiter Glo assay kit (Promega) according to the manufacturer’s protocol. The luminescence signal was recorded using TR717 microplate luminometer (Berthold) and WinGlow software (Berthold). The data was normalized to vehicle control and fitted to a curve using nonlinear regression method in GraphPad Prism 5 software (GraphPad Software).
Winglow software
Manufactured by Berthold Technologies
Sourced in Germany
WinGlow is a software application developed by Berthold Technologies for use with their laboratory equipment. The core function of WinGlow is to provide a platform for data acquisition, analysis, and visualization for various scientific applications.
Automatically generated - may contain errors
Lab products found in correlation
Mikrowin 2000 version 4, by Berthold Technologies (2 mentions)
Celltiter glo, by Promega (2 mentions)
White 96 well plate, by Greiner (1 mentions)
Prism 5, by GraphPad (1 mentions)
Penicillin streptomycin, by Corning (1 mentions)
Celltiter glo assay kit, by Promega (1 mentions)
Fetal bovine serum (fbs), by Avantor (1 mentions)
Trypsin, by Corning (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Corning (1 mentions)
Dulbecco s modified eagle s media, by Corning (1 mentions)
4 protocols using winglow software
1
Cytotoxicity Assay for HEK293T Cells
2
Cell Viability Assay in NIH/3T3 Cells
3
Cell Viability Assay in NIH/3T3 Cells
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Cell viability analysis was performed using CellTiter-Glo (Promega) according to the manufacturer’s instructions, in NIH/3T3 cells. The assay was performed after 24 h of compound treatment, and measurements were done in triplicates. The measured values were analysed with WinGlow Software, MikroWin 2000, Version 4.41 (Berthold Technologies) and normalized to control. Quantifications are based on at least three independent experiments.
4
Diagnostic Accuracy of TRAK Assay
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Results of the Bridge Assay were calculated as IU/l with WinGlow software (Berthold Technologies, Bad Wildbad, Germany). ROC analysis was used for evaluation of diagnostic accuracy. Intraund inter-assay CVs were calculated following NCCLS guideline EP5 with 2 replicates per run and one run per day for at least 30 days [28] . Calculations were performed with custom S-scripts for the statistical environment R 2.10 [29] . A grey zone was calculated using values with an inter-assay CV between 15 and ≤ 20 %. For the TRAK human assay, grey zone was defined by the manufacturer between 1.0 IU/l and 1.5 IU/l.
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