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GeneBLAzer™ CALCR -CRE-bla FreeStyle™ 293F Cells

The GeneBLAzer® CALCR-CRE-bla Freestyle293 cells contain the CALCR receptor isoform (CTR2/CT(a)/CTRI1-), accession # NM_001742.2, stably integrated into the CellSensor® CRE-bla Freestyle293 cell line. The CellSensor® CRE-bla Freestyle293 cells contain a beta-lactamase (bla) reporter gene under control of the cAMP (CRE) response element.

CALCR-CRE-bla Freestyle293 cells have been tested for variable assay conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time and functionally validated for Z' and EC50 concentrations of calcitonin. Additional testing data using alternate stimuli are also provided. These data are found in the Validation & Assay Performance Summary in the Manuals and Brochures section.

GeneBLAzer™ NPSR1-B-NFAT-bla CHO-K1 Cells

The GeneBLAzer® NPSR1-B-NFAT-bla CHO-K1 cells contain the human Neuropeptide S Receptor 1-Isoform B (NPSR1-B), (Accession # NP_997056) stably integrated into the CellSensor® NFAT-bla CHO-K1 cell line. CellSensor® NFAT-bla CHO-K1 cells (Cat #K1078) contain a beta-lactamase (bla) reporter gene under control of the NFAT response element.

The NPSR1-B-NFAT-bla CHO-K1 cells are functionally validated for Z'-factor and EC50 concentrations of Neuropeptide-S. In addition, NPSR1-B-NFAT-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary in the Manuals and Brochures section.

GeneBLAzer™ CRHR1-CRE-blaCHO-K1 Cells

GeneBLAzer® CRHR1-CRE-bla CHO-K1 cells contain the human Corticotropin Releasing Factor Receptor 1 (CRHR1) stably integrated into the CellSensor® CRE-bla CHO-K1 cell line. CellSensor® CRE-bla CHO-K1 cells (Cat. no. K1535) contain a beta-lactamase reporter gene under control of the Cyclic AMP Response Element (CRE) response element.

The GeneBLAzer® CRHR1-CRE-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of CRF. In addition, GeneBLAzer® CRHR1-CRE-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

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 revealed no major differences between them.

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GeneBLAzer™ MC2R-CRE-bla-CHO-K1 Cells

The GeneBLAzer® MC2R-CRE-bla-CHO-K1 cells contain the human melanocortin-2 receptor (MC2R), (Accession # NM_000529.1) and the Melanocortin-2 receptor accessory protein (MRAP), (Accession #NM_178817.3) stably integrated into the CellSensor® CRE-bla CHO-K1 cell line. CellSensor® CRE-bla CHO-K1 cells (Cat # K1129) contain a beta-lactamase (bla) reporter gene under control of the cAMP Response Element (CRE).

The MC2R-CRE-bla-CHOK1 cells are functionally validated for Z'-factor and EC50 concentrations of ACTH 1-24. In addition, MC2R-CRE-bla-CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary in the Manuals and Brochures section.

GeneBLAzer™ HCRTR2-Gα15-NFAT-bla CHO-K1 Cells

GeneBLAzer® HCRTR2-Gα15-NFAT-bla CHO-K1 cells contain the human Orexin Receptor B (HCRTR2) stably integrated into the GeneBLAzer® Gα15-NFAT-bla CHO-K1 cell line. GeneBLAzer® Gα15-NFAT-bla CHO-K1 cells (Cat. no. K1537) contain a beta-lactamase reporter gene under control of the Nuclear Factor of Activated T-cells (NFAT) response element and the Promiscuous G protein, Gα15.

The GeneBLAzer® HCRTR2-Gα15-NFAT-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of Orexin B. In addition, GeneBLAzer® HCRTR2-Gα15-NFAT-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

The HCRTR2 receptor has been detected in the brain as well as adrenal gland, gastrointestinal tract, and pancreas. The signaling cascades used by orexin receptors appear to be varied and cell type dependant. In CHO cells, calcium release is seen after agonist treatment. In hypothalamic neurons, the frequency of action potentials is increased. Administering either orexin peptide into the bloodstream of rats caused an increase in feeding behavior. The orexin system is also thought to regulate neuroendocrine function and the sleep wake cycle. HCRTR2 has been directly implicated in narcolepsy in animal studies.

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CellSensor™ TrkA-NFAT-bla CHO-K1 Cell Line

Neurotrophins (NGF, BDNF, NT-3, and NT-4) and their transmembrane receptors (TrkA, TrkB, TrkC, and P75NTR) play important roles in the regulation of neuronal and non-neuronal cell proliferation, differentiation, survival, and death. Neurotrophin signaling also mediates neuronal higher-order activities, such as learning, memory, and behavior. Alterations in neurotrophin levels and their receptors have been implicated in neurodegenerative disorders such as Alzheimer's disease and Huntington's disease, as well as psychiatric disorders. Neurotrophins propagate their signal by activating multiple signaling pathways. One of the signaling pathways of NGF (the ligand for TrkA) activates phospholipase C, releasing DAG and IP3, increasing downstream intracellular calcium, and activating protein kinase C. This promotes the translocation of the transcription factor, nuclear factor of activated T-cells (NFAT), from the cytosol into the nucleus, resulting in NFAT-dependent transcription.

The CellSensor® TrkA-NFAT-bla CHO-K1 Cell Line was engineered by integrating the human TrkA expression plasmid into the genome of existing CellSensor® NFAT-bla CHO-K1 Cell Line, which is engineered to express beta-lactamase under the control of NFAT. This cell line has been tested for assay performance under variable conditions, including DMSO concentration, cell number, stimulation time, substrate loading time, and in cryopreserved cells, and has been validated for Z´-factor and EC50 values under optimized conditions using NGF 2.5s.

Additional testing information using various small molecule inhibitors is also provided.

CellSensor™ GAS-bla ME-180 Cell Line

The CellSensor® GAS-bla ME-180 cell line contains a beta-lactamase reporter gene under control of the Gamma Activated Sequence (GAS) response element stably integrated into ME-180 cells. To construct this cell line, the GAS-bla construct was transduced into ME-180 cells by lentivirus. Flow cytometry was used to isolate cells responsive to Interferon Gamma (IFN-γ). This cell line is validated for DMSO tolerance, cell number, stimulation time, substrate loading time, as well as Z'and EC50 concentrations for IFN-&gamma. The CellSensor® GAS-bla ME-180 cell line is responsive to IFN-γ and can be used to probe the JAK/STAT signaling pathway. This cell line can be adapted for high-throughput screening of agonist or antagonist compound libraries. Candidate drugs can be tested for dose response against this cell line. Academic and non-profit customers, please inquire for special pricing.

Tango™ ADORA3-bla U2OS Cells

The Tango™ ADORA3-bla U2OS cells contain the human Adenosine A3 Receptor (ADORA3) linked to a TEV protease site and a Gal4-VP16 transcription factor stably integrated into the Tango™ GPCR-bla U2OS parental cell line. This parental cell line stably expresses a beta-arrestin⁄TEV protease fusion protein and the beta-lactamase (bla) reporter gene under the control of a UAS response element.

The Tango™ ADORA3-bla U2OS cells cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ ADORA3-bla U2OS cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary for each product.

GeneBLAzer™ D5-CRE-bla CHO-K1 Cells

The GeneBLAzer® D5-CRE-bla CHO-K1 cells contain the human Dopamine Receptor 5 (D5) 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 CRE response element.

The GeneBLAzer® D5-CRE-bla CHO-K1 cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, GeneBLAzer® D5-CRE-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary for each product.

GeneBLAzer™ VPAC2-CRE-bla CHO-K1 Cells

GeneBLAzer® VPAC2-CRE-bla CHO-K1 cells contain the human Vasoactive intestinal peptide/pituitary adenylate cyclase activating polypeptide receptor 2 (VPAC2) stably integrated into the CellSensor® CRE-bla CHO-K1 cell line. CellSensor® CRE-bla CHO-K1 cells (Cat. no. K1535) contain a beta-lactamase reporter gene under control of the Cyclic AMP Response Element (CRE) response element.

The GeneBLAzer® VPAC2-CRE-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of VIP. In addition, GeneBLAzer® VPAC2-CRE-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

VPAC2 has been implicated in a wide range of autoimmune and inflammation disorders, including Chronic obstructive pulmonary disease, Chronic Fatigue Syndrome. VPAC2 may also play a role in Type-2 diabetes.

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CellSensor™ Myc-bla HCT116 Cell Line

The CellSensor™Myc-bla HCT116 cell line contains a beta-lactamase reporter gene under the control of Myc binding sequences. The construct was transduced into HCT116 cells using a lentiviral system. HCT116 is a colon cancer cell line which expresses a mutated form of beta-catenin. This form of beta-catenin leads to the accumulation of beta-catenin and constitutive activation of downstream genes such as Myc. This cell line is a clonal population isolated by flow cytometry. It has been validated for cell plating density and DMSO tolerance. The signaling pathway has been validated using RNAi against c-Myc and ICG-001, an inhibitor of the wnt-beta-catenin pathway. The expression of the mutant beta-catenin in HCT116 cells results in constitutive activation of beta-lactamase in this CellSensor™line, which can be knocked down by ICG-001 (Figure 1) or Myc RNAi (Figure 2).

GeneBLAzer™ MC4R-CRE-bla CHO-K1 Cells

GeneBLAzer® MC4R-CRE-bla CHO-K1 cells contain the human Melanocortin Receptor 4 (MC4R) stably integrated into the CellSensor® CRE-bla CHO-K1 cell line. CellSensor® CRE-bla CHO-K1 cells (Cat. no. K1535) contain a beta-lactamase reporter gene under control of the Cyclic AMP Response Element (CRE) response element.

The GeneBLAzer® MC4R-CRE-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of α MSH In addition, GeneBLAzer® MC4R-CRE-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

MC4 receptors are expressed almost exclusively in the CNS and bind all of the melanocortin peptides, but only have high affinity for α-MSH and ACTH. The agouti peptide binds to the MC4R receptor with high affinity in in vitro tests but, being normally expressed only in skin, doesn't have access to the MC4R receptor under physiological conditions. The AGRP peptide is a high-affinity antagonist of the neural MC4R receptor. The MC4R receptor is involved in the regulation of energy homeostasis and erectile function.

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Tango™ SSTR5-bla U2OS Cells

The Tango™ SSTR5-bla U2OS cells contain the human Somatostatin Receptor 5 (SSTR5) linked to a TEV protease site and a Gal4-VP16 transcription factor stably integrated into the Tango™ GPCR-bla U2OS parental cell line. This parental cell line stably expresses a beta-arrestin⁄TEV protease fusion protein and the beta-lactamase (bla) reporter gene under the control of a UAS response element.

The Tango™ SSTR5-bla U2OS cells cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ SSTR5-bla U2OS cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary for each product.

GeneBLAzer™ VDR-UAS-bla HEK 293T Cells

VDR-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human Vitamin D receptor (VDR) fused to the DNAbinding domain of GAL4 stably integrated in the GeneBLAzer®UAS-bla HEK 293T cell line. GeneBLAzer®UAS-bla HEK 293T cells stably express a beta-lactamase reporter gene under the transcriptional control of an upstream activator sequence (UAS). When an agonist binds to the LBD of the GAL4 (DBD)-VDR (LBD) fusion protein, the protein binds to the UAS, resulting in expression of beta-lactamase. 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. VDR-UAS-bla HEK 293T cells are functionally validated for Z' and EC50 concentrations of Calcitriol (1α, 25-dihydroxyvitamin D3). In addition, VDR-UAS-bla HEK 293T cells have been tested for assay performance under variable conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time.

Tango™ EDG8-bla U2OS Cells

The Tango™ EDG8-bla U2OS cells contain the human Endothelial Differentiation Gene 8 (EDG8) linked to a TEV protease site and a Gal4-VP16 transcription factor stably integrated into the Tango™ GPCR-bla U2OS parental cell line. This parental cell line stably expresses a beta-arrestin/TEV protease fusion protein and the beta-lactamase (bla) reporter gene under the control of a UAS response element.

The Tango™ EDG8-bla U2OS cells are functionally validated for Z' and EC50 concentrations of S1P. In addition, Tango™ EDG8-bla U2OS cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

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