For determining cell viability after CBX treatment, co-cultures of GS-293 and FSHR-293 were preincubated with different concentrations of CBX (25, 50, 75 and 100 μM) for 2h in assay medium. Assay medium was then discarded, followed by washing with PBS. Cell viability was then assessed by CellTiter AQueous non-radioactive cell proliferation assay (Promega) following manufacturer’s instructions. Briefly, cells were incubated in DMEM complete medium (without phenol red) containing MTS/PMS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium/phenazine methosulfate] solution for 2h (37°C, 5% CO2). Absorbance was then read at 490 nm, which provides a measure of cellular viability. One-way analysis of variance (ANOVA) was used to determine statistical differences among different samples.
Memerald
MEmerald is a lab equipment product. It is a fluorescent protein that can be used for imaging and visualization applications in biological research.
Lab products found in correlation
4 protocols using memerald
Connexin-32 Overexpression and Cell Viability
For determining cell viability after CBX treatment, co-cultures of GS-293 and FSHR-293 were preincubated with different concentrations of CBX (25, 50, 75 and 100 μM) for 2h in assay medium. Assay medium was then discarded, followed by washing with PBS. Cell viability was then assessed by CellTiter AQueous non-radioactive cell proliferation assay (Promega) following manufacturer’s instructions. Briefly, cells were incubated in DMEM complete medium (without phenol red) containing MTS/PMS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium/phenazine methosulfate] solution for 2h (37°C, 5% CO2). Absorbance was then read at 490 nm, which provides a measure of cellular viability. One-way analysis of variance (ANOVA) was used to determine statistical differences among different samples.
Fluorescent Protein Transfection Protocol
Fluorescent Protein-Tagged Receptor Constructs
The mCherry-VEGFR2 pcDNA3.1 construct contained VEGFR2 cDNA inserted into the KpnI site followed by a linker, and in frame with the mCherry sequence. The linker sequence was GGTGGGAGTGGAGGTGGGAGTGGA. The wild type receptor construct was used as a template for the mutants. The KpnI-AgeI receptor fragment was replaced with the corresponding mutant fragments that were obtained from previously generated cDNAs [22 (link)]. All constructs were sequence verified.
Plasmid Repository for Cellular Imaging
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