Thermo Scientific™ CellSensor™ HRE-blaHCT-116 Cell Line

Description

• Hypoxia is an almost universal hallmark of solid tumors
• 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)
• Under conditions of normal oxygenation, the regulated HIF-1α is hydroxylated and degraded by the proteasome system
• 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
• The CellSensor™ HRE-bla HCT-116 Cell Line contains a beta-lactamase reporter gene under control of the HRE stably integrated into HCT-116 cells
• 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 hr
• with agonist and 0.5% DMSO and then combined with LiveBLAzer™-FRET B/G Substrate (CCF4 - AM) for 2 hr
• 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
• CellSensor™ HRE-bla HCT-116 cells were stimulated with Cobalt Chloride (CoCl 2 ) over the indicated concentration range in a 384-well format
• Cells were incubated for 24 hr
• with agonist and 0.5% DMSO and then combined with LiveBLAzer™-FRET B/G Substrate (CCF4 - AM) for 2 hr
• Fluorescence emission values at 460 nm and 530 nm were obtained using a standard fluorescence plate reader
• Response ratios were plotted against the indicated concentrations of CoCl2
• Academic and non-profit customers, please inquire for special pricing.