GeneBLAzer™ CRHR1 CHO-K1 DA Assay Kit

GeneBLAzer™ CRHR1 CHO-K1 DA (Division Arrested) cells and CRHR1-CRE-bla CHO-K1 cells contain the human Corticotropin Releasing Factor 1 (CRHR1) receptor stably integrated into the CellSensor™ CRE-bla CHO-K1 cell line. CellSensor™ CRE-bla CHO-K1 cells (Cat. no.K1129) contain a beta-lactamase reporter gene under control of the Cyclic AMP Response Element (CRE). Division Arrested (DA) cells are available in two configurations- an Assay Kit (which includes cells and sufficient substrate to analyze 1 x 384-well plate), and a tube of cells sufficient to analyze 10 x 384-well plates.

DA cells are irreversibly division arrested using a low-dose treatment of Mitomycin-C, and have no apparent toxicity or change in cellular signal transduction. Both CRHR1 CHO-K1 DA cells and CRHR1-CRE-bla CHO-K1 cells are functionally validated for Z’-factor and EC₅₀ concentrations of corticotropin releasing factor (CRF), (Figure 1). In addition, CRHR1-CRE-bla CHO-K1 cells have been tested for assay performance under variable conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time (data available upon request). Additional testing data using alternate stimuli are also available.

Corticotropin releasing factor is a 41-amino acid peptide that plays a role in the integration of autonomic, neuroendocrine, and behavioral responses to stress. These effects are mediated through two receptor families, CRHR1 and CRHR2. While CRF was originally isolated from the hypothalamus, where it was shown to be the primary neuroregulator mediating the hypothalamic-pituitary-adrenocortical stress axis, it has since been found to be widely distributed outside the hypothalamus throughout the central nervous system. Presently, there are five distinct targets for CRF with unique pharmacology and localization. These have been placed into three distinct classes, two of which are the G-protein-coupled receptors CRF1 (CRHR1) and CRF2 (CRHR2). Three functional splice variants have been identified for the mammalian CRHR2 receptor, although pharmacological characterization of these splice variants has revealed no major differences between them.

Storage:
GeneBLAzer™ CRHR1 CHO-K1 DA Cells and CRHR1-CRE-bla CHO-K1 Cells are shipped on dry ice. Store in liquid nitrogen immediately upon receipt or thaw for immediate use.


Contents:
GeneBLAzer™ CRHR1 CHO-K1 DA Assay Kit
Each system contains sufficient division arrested (DA) cells & substrate to assay 1 x 384-well plate. (Other materials are required separately; please refer to the protocol). Includes:
•CRHR1 CHO-K1 DA cells
•LiveBLAzer™-FRET B/G Loading Kit, 70μg
•Solution D, 1ml
•2 protocols
•Certificate of Analysis
GeneBLAzer™ CRHR1 CHO-K1 DA cells
Each system contains sufficient division arrested (DA) cells to assay 10 x 384-well plates (LiveBLAzer™-FRET B/G Loading Kit, Solution D and other materials are required separately; please refer to the protocol).
Includes:
•CRHR1 CHO-K1 DA cells
•1 protocol
•Certificate of Analysis
GeneBLAzer™ CRHR1-CRE-bla CHO-K1 cells
(LiveBLAzer™-FRET B/G Loading Kit, Solution D and other materials are required separately; please refer to the protocol).
Includes:
•CRHR1-CRE-bla CHO-K1 cells
•1 protocol
•Certificate of Analysis