CellSensor™ HRE-bla HCT-116 Cell Line

Description

• Adaptation to hypoxia is critical for tumor survival and growth, and is mediated largely by transcriptional activation of genes that facilitate short - (e.g., glucose transport) and long - term (e.g., angiogenesis) adaptive mechanisms.
• This coordinated homeostatic response is mediated in large part through the activation of the heterodimeric transcription factor hypoxia - inducible factor 1 (HIF - 1).
• As oxygen becomes rate limiting, hydroxylation diminishes and HIF - 1α accumulates and heterodimerizes with the constitutively present α - subunit.
• The binding of this complex to the cognate hypoxia - response element (HRE) results in transcriptional activation of genes containing such elements within promoter or enhancer elements.
• This cell line is validated for EC50 and Z'; - factor under optimized conditions using deferoxamine (DFO) and Cobalt Chloride.
• This cell line has also been tested under various experimental conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time.
• CellSensor ™ HRE-bla HCT - 116 cells were stimulated with deferoxamine (DFO) over the indicated concentration range in a 384 - well format.
• Cells were incubated for 24 hrs with agonist and 0.5% DMSO and then combined with LiveBLAzer™ - FRET B/G Substrate (CCF4 - AM) for 2 hrs.
• Fluorescence emission values at 460 nm and 530 nm were obtained using a standard fluorescence plate reader.
• The Response Ratios were plotted against the indicated concentrations of DFO.
• Response ratios were plotted against the indicated concentrations of CoCl2.