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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.

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.

For Academic pricing please login or contact us at discoverysciences@invitrogen.com.

BacMam ERK2 [pThr185/pTyr187] Cellular Assay Kit (Invitrogen™)

The BacMam ERK2 [pThr185⁄pTyr187] Cellular Assay Kit is a high-throughput screening (HTS) compatible cellular immunoassay for the measuring of phosphorylation of GFP-ERK2. The BacMam ERK2 Cellular Assay Kit can be used with numerous transformed cell lines, primary cells, and stem cells, and allows for the interrogation of ERK2, either as a kinase or GPCR readout.

• Choose cell types that are relevant to your needs
• Get results in weeks instead of months
• Perform Assays on Your Schedule

Use Cell Types Relevant to Your Needs
Understanding signaling in physiologically relevant cell backgrounds is extremely important when predicting the real effects of compounds in humans prior to clinical trials. With the BacMam ERK2 [pThr185⁄pTyr187] Cellular Assay Kit you can choose the appropriate cell type for your research needs, enabling you to design your own experiments, and to reach your research goals faster.

BacMam ERK2 Reagent Compatible Cells
The BacMam reagent included in the kit has been used to successfully transduce numerous transformed cell lines (e.g. U2-OS, HEK 293, and HeLa), primary cells (e.g., fibroblasts, hepatocytes, cardiovascular cells and epithelial cells), and stem cells (e.g. neuronal and mesenchymal) at high transduction efficiency and low levels of cytotoxicity. However, cells of hematopoietic origin show consistently poor transduction by BacMam technology, and are not recommended for use with the current technology.

Develop Assays Faster
The BacMam platform enables transient target expression allowing for reduced time to an HTS ready assay versus traditional stable cell line creation. Traditional stable cell line development requires months of culturing, optimization, and cloning of your cell line. This requires considerable time and resources. With BacMam ERK2 a user can get results in weeks versus months.

Cells Ready When You Are
BacMam Erk2 Reagent is ideal to transduce large quantities of cells in batch mode; transduced cells can be stored frozen in aliquots for later use. This saves you the delays associated with culturing cells. The cells are ready whenever you are. Cells can be assayed typically within hours of thawing, providing assay-ready cells when you need them, with no loss of activity.

For Research Use Only. Not intended for animal or human therapeutic or diagnostic use.

GeneBLAzer™ PAC1 CHO-K1 DA Assay Kit

The GeneBLAzer® PAC1-CRE-bla CHO-K1 cells contain the human Adenylate Cyclase Activating Polypeptide 1 Receptor 1 (ADCYAP1R⁄PAC1) 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 PAC1-CRE-bla CHO-K1 cells are functionally validated for Z'-factor and EC50 concentrations of established ligands. In addition, PAC1-CRE-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary. Division Arrested cells are irreversibly division arrested using a low-dose treatment of Mitomycin C, and have no apparent toxicity or change in cellular signal transduction.

Tango™ EDG2-bla U2OS Cells

The Tango™ EDG2-bla U2OS cells contain the human Endothelial Differentiation Gene 2 (EDG2) 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™ EDG2-bla U2OS cells are functionally validated for Z' and EC50 concentrations of LPA (18: 1). In addition, Tango™ EDG2-bla U2OS cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

For Academic pricing please login or contact us at discoverysciences@invitrogen.com.

Tango™ EDG1-bla U2OS Cells

The Tango™ EDG1-bla U2OS cells contain the human Endothelial Differentiation, Sphingolipid G-protein-coupled Receptor, 1 (EDG1) 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™ EDG1-bla U2OS cells cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ EDG1-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™ Gα15-NFAT-bla HEK 293T Cells

The GeneBLAzer® Gα15-NFAT-bla HEK 293T cell line contains the Gα protein Gα15 stably integrated into the CellSensor™ NFAT-bla HEK 293T parental cell line. NFAT-bla HEK 293T cells contain a beta-lactamase reporter gene under control of an NFAT response element stably integrated into HEK 293T cells. The Gα15-NFAT-bla HEK 293T master cell line is designed for the construction of Gi/o-coupled GPCR cell-based assays (Figure 1).

BacMam Histone H3 [AcLys9] Cellular Assay Kit

The combination of baculovirus-mediated gene delivery (BacMam) with LanthaScreen® Cellular Assay technology enables a platform for the analysis of specific posttranslational modifications of histones. BacMam provides a convenient genetic delivery tool for a GFP-Histone H3 fusion protein in the cell line of interest. This kit describes an HTS-compatible cellular immunoassay measuring acetylation of GFP-Histone H3 at Lys9.

GeneBLAzer™ NPSR1-A CHO-K1 DA Assay Kit

The GeneBLAzer® NPSR1-A CHO-K1 DA (Division Arrested) cells contain the human Neuropeptide S Receptor 1-Isoform A stably integrated into the CellSensor® NFAT-bla CHO-K1 cell line. CellSensor® NFAT-bla CHO-K1 cells (Cat # K1129) contain a beta-lactamase (bla) reporter gene under control of the nuclear factor of activated T-cell (NFAT) response element.

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

Tango™ CXCR2-bla U2OS Cells

The Tango™ CXCR2-bla U2OS cells contain the Chemokine (C-X-C motif) Receptor 2 (CXCR2) 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™ CXCR2-bla U2OS cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ CXCR2-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.

CellSensor™ CRE-bla HEK 293T Cell Line

The CellSensor® CRE-bla HEK 293T cell line contains the b-lactamase reporter gene under control of the cAMP response element (CRE). To obtain the cell line, the CRE-bla construct was transduced into HEK 293T cells by lentivirus. Subsequent flow cytometry was used to isolate clones responsive to forskolin stimulation. The CellSensor® CRE-bla HEK 293T cell line can be used to build specific G-protein coupled receptor (GPCR) assays or to detect changes in intracellular cAMP levels. Academic and non-profit customers, please inquire for special pricing.

BacMam Akt [pSer473] Cellular Assay Kit

The combination of baculovirus-mediated gene delivery (BacMam) with LanthaScreen® Cellular Assay technology enables a platform for the analysis of specific posttranslational modifications of various substrates including AKT. BacMam provides a convenient genetic delivery tool for a GFP-AKT fusion protein in the cell line of interest. This kit describes an HTS-compatible cellular immunoassay measuring phosphorylation of GFP-AKT at Ser473.

Tango™ EDG6-bla U2OS Cells

The Tango™ EDG6-bla U2OS cells contain the human Endothelial Differentiation, Lysophosphatidic Acid G-protein-coupled Receptor, 6 (EDG6) 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™ EDG6-bla U2OS cells cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ EDG6-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.

CellSensor™ irf1-bla HEL Cell Line

Jak/Stat signaling pathways play essential roles in the cellular responses to distinct cytokines. One of Jak/Stat pathways, Jak2/Stat5, mediates cell proliferation in response to Interleukin-3 (IL-3), prolactin, erythropoietin (Epo), and granulocyte-macrophage colony stimulating factor (GM-CSF). JAK2 gene knock-out causes embryonic lethality due to defective erythropoiesis, suggesting the Jak2/Stat5 pathway plays important role in red blood cell formation. Recent discovery of activating mutation in JAK2 (V617F) present in high percentage of myeloproliferative disease (MPD) patients suggests Jak2/Stat5 pathway to be the potential therapeutic target for certain forms of MPD. The activated transcription factor Stat5 dimers recognize and bind to a specific palindromic DNA sequence found in the promoter region of β-casein, interferon regulatory factor-1 (irf-1) and a number of other genes. The CellSensor® irf1-bla HEL cell line contains a beta-lactamase reporter gene under control of the interferon regulatory factor-1 (irf1) response element stably integrated into HEL cells. HEL cells are a human erythroleukemia cell line that is growth factor independent and contains a endogenous homozygous JAK2V617F mutation. This cell line validated for IC50 and Z'-Factor under optimized conditions using Jak Inhibitor 1 (Figure 1). This cell line has also been tested under variable experimental conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time. Academic and non-profit customers, please inquire for special pricing.

Tango™ OPRM1-bla U2OS Cells

The Tango™ OPRM1-bla U2OS cells contain the human Opioid Receptor Mu 1 (OPRM1) 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™ OPRM1-bla U2OS cells have been functionally validated for Z' factor and EC50 concentrations of DAMGO. In addition, Tango™ OPRM1-bla U2OS 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™ Gqi5-NFAT-bla FreeStyle™ 293F Cells

GeneBLAzer® Gqi5-NFAT-bla FreeStyle™ 293F cells contain the chimeric G-protein Gqi5 stably integrated into the CellSensor® NFAT-blaFreeStyle™ 293F cell line. CellSensor® NFAT-bla FreeStyle™ 293F cells contain a beta-lactamase reporter gene under control of a stably integrated NFAT response element. Gqo5-NFAT-bla FreeStyle™ 293F is a parental cell line designed to be used for the construction of Gi/o-coupled GPCR beta-lactamase cellular assays. The Gi/o receptor of interest should be stably expressed in the cell line by expression from the selectable vector pcDNA™6.2-hygro-DEST (Cat. no. K1233). Gqi5-NFAT-bla FreeStyle™ 293F cells have been shown to respond to the agonist of a transiently expressed D2 receptor.

GeneBLAzer™ CysLT2-NFAT-bla CHO-K1 Cells

GeneBLAzer® CysLT2-NFAT-bla CHO-K1 cells contain the human Cysteinyl receptor (CysLT2) stably integrated into the CellSensor® NFAT-bla CHO-K1 cell line. CellSensor® NFAT-bla CHO-K1 cells (Cat. no. K1534) contain a beta-lactamase reporter gene under control of the Nuclear Factor of Activated T-cells (NFAT) response element.

The GeneBLAzer® CysLT2-NFAT-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of LTD4. In addition, GeneBLAzer® CysLT2-NFAT-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

Cysteinyl leukotrienes play a role in the patho-physiology of numerous inflammatory disorders, including chronic asthmatic inflammation. The cysteine leukotriene receptors, CysLT1 and CysLT2, are thus primary targets for anti-asthma drugs. A number of cysteinyl leukotrienes (including LTC4, LTD4, and LTE4) have been identified, and are known to function as inflammatory mediators as well as being potent smooth muscle contractile agents.

Leukotriene receptor antagonists (LTRAs) reduce the effect of allergic inflammation and athsma by blocking cysteine leukotriene activity. Several CysLT antagonists, such as montelukast, zafirlucast, and pranlukast, are currently used in the treatment of asthma.

For Academic pricing please login or contact us at discoverysciences@invitrogen.com.

Tango™ CCR1-bla U2OS DA Assay Kit

The Tango™ CCR1-bla U2OS DA cells contain the human Chemokine (C-C Motif) Receptor 1 (CCR1) 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™ CCR1-bla U2OS DA cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ CCR1-bla U2OS DA cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary for each product. Division Arrested cells are irreversibly division arrested using a low-dose treatment of Mitomycin C, and have no apparent toxicity or change in cellular signal transduction.

BacMam Akt [pThr308] Cellular Assay Kit

The combination of baculovirus-mediated gene delivery (BacMam) with LanthaScreen® Cellular Assay technology enables a platform for the analysis of specific posttranslational modifications of various substrates including AKT. BacMam provides a convenient genetic delivery tool for a GFP-AKT fusion protein in the cell line of interest. This kit describes an HTS-compatible cellular immunoassay measuring phosphorylation of GFP-AKT at Thr308.

Tango™ SSTR1-bla U2OS Cells

The Tango™ SSTR1-bla U2OS cells contain the human Somatostatin Receptor 1 (SSTR1) 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™ SSTR1-bla U2OS cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ SSTR1-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™ CCKAR HEK 293T DA Assay Kit

The GeneBLAzer® CCKAR-NFAT-bla CHO-K1 cells contain the human Cholecystokinin A Receptor (CCKAR) stably integrated into the GeneBLAzer® NFAT-bla CHO-K1 cell line. GeneBLAzer® NFAT-bla CHO-K1 (Cat # K1447) contains a beta-lactamase (bla) reporter gene under control of a NFAT response element stably integrated into CHO-K1 cells.

The CCKAR-NFAT-bla CHO-K1 cells are functionally validated for Z'-factor and EC50 concentrations using established ligands. In addition, CCKAR-NFAT-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary. Division Arrested cells are irreversibly division arrested using a low-dose treatment of Mitomycin C, and have no apparent toxicity or change in cellular signal transduction.

CellSensor™ SIE-bla ME-180 Cell Line

The CellSensor® SIE-bla ME-180 cell line contains a beta-lactamase reporter gene under control of the Sis-Inducible Element (SIE) stably integrated into ME-180 cells. To construct this cell line, the SIE-bla construct was transduced into ME-180 cells by lentivirus. Flow cytometry was used to isolate cells responsive to Interleukin-6 (IL-6). This cell line is validated for DMSO tolerance, cell number, stimulation time, substrate loading time, as well as Z' and EC50 concentrations of IL-6. The CellSensor® SIE-bla ME-180 cell line is responsive to IL-6, OSM and IFNγ 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 also be tested for dose response against this cell line. Academic and non-profit customers, please inquire for special pricing.

GeneBLAzer™ OXTR-Gqo5-NFAT-bla CHO-K1 Cells

GeneBLAzer® OXTR-Gqo5-NFAT-bla CHO-K1 cells contain the human Oxytocin Receptor (OXTR), (Accession # NM_002531) stably integrated into the GeneBLAzer® Gqo5-NFAT-bla CHO-k1 cell line. GeneBLAzer® Gqo5-NFAT-bla CHO-k1 cells (Cat. no.K1536) contain a beta-lactamase reporter gene under control of the NFAT. The GeneBLAzer® OXTR-Gqo5-NFAT-bla CHO-K1 cells are functionally validated for Z’-factor and EC50 concentrations of Oxytocin.

Tango™ HRH3-bla U2OS DA Assay Kit

The Tango™ HRH3-bla U2OS DA (Division Arrested) cells contain the human Histamine Receptor H3 (HRH3) 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™ HRH3-bla U2OS cells and the Tango™ HRH3-bla U2OS DA cells have been functionally validated for Z' factor and EC50 concentrations of Methylhistamine. In addition, Tango™ HRH3-bla U2OS 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. 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.

GeneBLAzer™ CNR1-Gα15-NFAT-bla CHO-K1 Cells

GeneBLAzer® CNR1-Gα15-NFAT-bla CHO-K1 cells contain the human Cannabinoid Receptor 1 (CNR1) 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® CNR1-Gα15-NFAT-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of CP-55,940. In addition, GeneBLAzer® CNR1-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 cannabinoid receptors are a family of GPCRs named after their endogenous ligand Delta9-tetrahydrocannabinol (Delta9-THC). Delta9-THC is the psychoactive component of Cannabis sativa (marijuana). CNR1 is present in many tissues throughout the body. The highest concentrations of cannabinoid receptors are in the neurons of the brain, the hippocampus, cerebral cortex, basal ganglia, and the cerebellum. Low-level expression occurs in the lung, testis, and uterus, as well as vascular tissue. Delta9-THC and CP55,940 are endogenous agonists of the CNR1 pathway. Several Delta9-THC and CP55,940 analogs have been synthesized, including HU 210 and WIN 55,212-2. The CNR1 and CNR2 receptors have equal affinity for cannabinoid agonists, but several antagonists that are subtype selective have been synthesized. Examples of these antagonists are AM251, LY-320135, and SR 141716A.

For Academic pricing please login or contact us at discoverysciences@invitrogen.com.

Tango™ CXCR6-bla U2OS Cells

The Tango™ CXCR6-bla U2OS cells contain the human Chemokine (C-X-C motif) receptor 6 (CXCR6) 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™ CXCR6-bla U2OS cells have been functionally validated for Z' factor and EC50 concentrations of CXCL16. These data are found in the Validation & Assay Performance Summary in the Manuals and Brochures section.

Tango™ HTR1D-bla U2OS Cells

The Tango™ HTR1D-bla U2OS cells contain the human 5-Hydroxytryptamine (Serotonin) Receptor 1D (HTR1D) 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™ HTR1D-bla U2OS cells cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ HTR1D-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.

Tango™ MLNR-bla U2OS Cells

Tango™ MLNR-bla U2OS cells contain the human Motilin Receptor (MLNR) 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 reporter gene under the control of a UAS response element. The Tango™ MLNR-bla U2OS cells have been functionally validated for Z’ factor and EC50 concentrations of Motilin.

Tango™ GLP2R-bla U2OS Cells

The Tango™ GLP2R-bla U2OS cells contain the human Glucagon-Like Peptide 2 Receptor (GLP2R) 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™ GLP2R-bla U2OS cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ GLP2R-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.

Tango™ AGTRL1-bla U2OS Cells

The Tango™ AGTRL1-bla U2OS cells contain the human Angtiotension-Like Receptor 1 (AGTRL1) 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™ AGTRL1-bla U2OS cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™AGTRL1-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™ GPR10-NFAT-bla CHO-K1 Cells

GeneBLAzer® GPR10 CRE-bla CHO-K1 cells contain the human Prolactin Releasing Peptide (GPR10) 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® GPR10 CRE-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of PrRP20. In addition, GeneBLAzer® GPR10 CRE-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

The GPR10 receptor has been implicated in a variety of metabolic disorders, including lactation, the regulation of food intake, and other physiological functions.

For Academic pricing please login or contact us at discoverysciences@invitrogen.com.

GeneBLAzer™ P2RY2-NFAT-bla CHO-K1 Cells

GeneBLAzer® P2RY2-NFAT-bla CHO-K1 cells contain the human Purinergic receptor P2, G protein-coupled, 2 (P2RY2) stably integrated into the CellSensor® NFAT-bla CHO-K1 cell line. CellSensor® NFAT-bla CHO-K1 cells (Cat. no. K1534) contain a beta-lactamase reporter gene under control of the Nuclear Factor of Activated T-cells (NFAT) response element.

The GeneBLAzer® P2RY2-NFAT-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of ATP. In addition, GeneBLAzer® P2RY2-NFAT-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

P2RY2 expression has been identified in smooth muscle, skeletal muscle, heart, spleen, lymphocytes, macrophages, bone marrow, lung, intestine, placenta, brain, kidney, and liver. P2RY2 has been implicated in the induction of chloride secretion in airway epithelial tissue, increases in coronary blood flow by vasodilation, and epidermal homeostasis.

For Academic pricing please login or contact us at discoverysciences@invitrogen.com.

CellSensor™ irf1-bla TF-1 Cell Line

Jak/Stat signaling pathways play essential roles in cellular responses to distinct cytokines. One of the Jak/Stat pathways, Jak2/Stat5, mediates cell proliferation in response to Interleukin - 3 (IL - 3), prolactin, erythropoietin (Epo), and granulocyte - macrophage colony stimulating factor (GM - CSF). JAK2 gene knockout causes embryonic lethality due to defective erythropoiesis, suggesting that the Jak2/Stat5 pathway plays an important role in red blood cell formation. The recent discovery of an activating mutation in JAK2 (V617F) present in a high percentage of myeloproliferative disease (MPD) patients suggests that the Jak2/Stat5 pathway may be a potential therapeutic target for certain forms of MPD. The activated Stat5 transcription factor recognizes and binds to a specific palindromic DNA sequence found in the promoter region of β - casein, interferon regulatory factor - 1 (irf - 1), and a number of other genes. The CellSensor® irf1 - bla TF1 Cell Line contains a beta - lactamase reporter gene under control of the irf - 1 response element stably integrated into TF1 cells. TF1 cells are a human erythroleukemia cell line that is growth - dependent on GM - CSF and have an intact GM - CSF - JAK2 - STAT5 pathway. This cell line is validated for EC50 and Z’ - factor using GM - CSF. This cell line has also been tested under variable experimental conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time. Responsiveness to EPO and Jak Inhibitor 1 was also tested (Figures 2 and 3). CellSensor® irf1 - bla TF1 cells were stimulated in triplicate with GM - CSF over the indicated concentration range in a 384 - well format. Cells were incubated for 5 hrs with agonist and 0.5% DMSO and then combined with LiveBLAzer™ - FRET B/G Substrate (CCF4 - AM) for 2.5 hrs. Fluorescence emission values at 460 nm and 530 nm were obtained using a standard fluorescence plate reader. The 460/530 ratios were plotted for each replicate against the indicated concentrations of GM - CSF. CellSensor® irf1 - bla TF1 cells were stimulated with Epo over the indicated concentration range in a 384 - well format. Cells were incubated for 5 hrs with agonist and 0.5% DMSO and then combined with LiveBLAzer™ - FRET B/G Substrate (CCF4 - AM) for 2.5 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 Epo. CellSensor® irf1 - bla TF1 cells were treated with Jak Inhibitor 1 over the indicated concentration range in a 384 - well format. Cells were then stimulated with GM - CSF or EPO for 5 hrs in 0.5% DMSO and then combined with LiveBLAzer™ - FRET B/G Substrate (CCF4 - AM) for 2.5 hrs. Fluorescence emission values at 460 nm and 530 nm were obtained using a standard fluorescence plate reader. These values were converted to percent inhibition relative to a set of controls (unstimulated cells and EC80 of GM - CSF or EPO treated cells) and plotted against the indicated concentrations of Jak Inhibitor 1. Academic and non-profit customers, please inquire for special pricing.

Tango™ CCR1-bla U2OS Cells

The Tango™ CCR1 -bla U2OS cells contain the human Chemokine (C-C Motif) Receptor 1 (CCR1) 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™ CCR1-bla U2OS cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ CCR1-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™ ADORA2A CHO-K1 DA Assay Kit

This cell line is available as dividing cells by requesting a quote for K1245 from dividingcells@invitrogen.com

GeneBLAzer® ADORA2A CHO-K1 DA (Division Arrested) cells and ADORA2A-CRE-bla CHO-K1 cells contain the human Adenosine A2a (ADORA2A) 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 ADORA2A CHO-K1 DA cells and ADORA2A-CRE-bla CHO-K1 cells are functionally validated for Z’-factor and EC50 concentrations of 5’-adenosine (NECA); (Figure 1). In addition, ADORA2A-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.

The Adenosine A2a Receptor (ADORA2A) has been shown to have many roles throughout the body. ADORA2A is involved in pain and inflammation regulation, inhibition of platelet aggregation, blood pressure regulation and contributes to ischemic brain damage. Selective antagonists of the ADORA2A pathway are being tested as adjuvants to dopaminergic drugs to treat Parkinson’s disease and schizophrenia.


Storage:

GeneBLAzer® ADORA2A CHO-K1 DA Cells and ADORA2A-CRE-bla CHO-K1 Cells are shipped on dry ice. Store in liquid nitrogen immediately upon receipt or thaw for immediate use.


Contents:
GeneBLAzer® ADORA2A 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:
•ADORA2A CHO-K1 DA cells
•LiveBLAzer™-FRET B/G Loading Kit, 70µg
•Solution D, 1ml
•2 protocols
•Certificate of Analysis
GeneBLAzer® ADORA2A 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:
•ADORA2A CHO-K1 DA cells
•1 protocol
•Certificate of Analysis
GeneBLAzer® ADORA2A-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:
•ADORA2A-CRE-bla CHO-K1 cells
•1 protocol
•Certificate of Analysis

GeneBLAzer™ TR beta DA Assay Kit

GeneBLAzer®TR beta DA(Division Arrested) cells and TR beta-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human Thyroid hormone receptor beta (TR beta) fused to the DNA-binding domain of GAL4 stably integrated in the GeneBLAzer®UASbla HEK 293T cell line. GeneBLAzer®UAS-bla HEK 293T cells stably express a betalactamase reporter gene under the transcriptional control of an upstream activator sequence (UAS). When an agonist binds to the LBD of the GAL4 (DBD)-TR beta (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. 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 TR beta DA cells and TR beta-UAS-bla HEK 293T cells are functionally validated for Z' and EC50 concentrations of T3 Thryoid hormone. In addition, TR beta-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 (data available upon request). Additional testing data using alternate stimuli are also available.

GeneBLAzer™ M1 CHO-K1 DA Assay Kit

This cell line is available as dividing cells by requesting a quote for K1230 from dividingcells@invitrogen.com

GeneBLAzer® M1 CHO-K1 DA(Division Arrested) cells and M1-NFAT-bla CHO-K1 cells contain the human Acetylcholine (muscarinic) subtype 1 receptor (M1) stably integrated into the CellSensor® NFAT-bla CHO-K1 cell line. CellSensor® NFAT-bla CHO-K1 cells (Cat. no.K1078) contain a beta-lactamase reporter gene under control of the Nuclear Factor of Activated T-cells (NFAT) response element. 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 M1 CHO-K1 DA cells and M1-NFAT-bla CHO-K1 cells are functionally validated for Z’-factor and EC50 concentrations of carbachol, (Figure 1). In addition, M1-NFAT-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.

The Musacarinic 1(M1) Receptor is localized principally in the cortical and hippocampal regions of the brain and has been implicated in various neurological conditions including depression, anxiety, and sleep disorders.


Storage:

GeneBLAzer® M1 CHO-K1 DA Cells and M3-NFAT-bla CHO-K1 Cells are shipped on dry ice. Store in liquid nitrogen immediately upon receipt or thaw for immediate use.


Contents:
GeneBLAzer® M1 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:
•M1 CHO-K1 DA cells
•LiveBLAzer™-FRET B/G Loading Kit, 70µg
•Solution D, 1ml
•2 protocols
•Certificate of Analysis
GeneBLAzer® M1 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:
•M1 CHO-K1 DA cells
•1 protocol
•Certificate of Analysis

GeneBLAzer® M1-NFAT-bla CHO-K1 cells
(LiveBLAzer™-FRET B/G Loading Kit, Solution D and other materials are required separately; please refer to the protocol).
Includes:
•M1-NFAT-bla CHO-K1 cells
•1 protocol
•Certificate of Analysis

CellSensor™ SIE-bla HEK293T Cell Line

The CellSensor® SIE-bla HEK 293T cell line contains a beta-lactamase reporter gene under control of the SIE response element stably integrated into HEK 293T cells. The cell line was created through sorting by FACS of cells responsive to stimulation of the SIE pathway with interleukin 6 (IL-6). The cell line was validated for DMSO tolerance, incubation time with stimulant, and substrate loading conditions. The CellSensor® SIE-bla HEK 293T cell line responds to agonist treatment as expected from literature and can be adapted for high throughput screening for agonists or antagonists of the SIE pathway with compound libraries. Candidate drugs can also be tested for dose response against this cell line. Academic and non-profit customers, please inquire for special pricing.

GeneBLAzer™ PPAR alpha UAS-bla HEK 293T Cellular Assay Kit

The GeneBLAzer® PPAR alpha-UAS-bla HEK 293T Cellular Assay provides an accurate, sensitive, and easy-to-use method for monitoring the cellular response of PPAR alpha to drug candidates or other stimuli.

Features of the The GeneBLAzer® PPAR alpha-UAS-bla HEK 293T Cellular Assay:
Convenience—plate cells and perform assay without additional cell culture steps
Low Noise—the kit uses a Fluorescence Resonance Energy Transfer (FRET)-based ratiometric readout for minimum noise and error
Low Background—the GAL4 fusion prevents any background from endogenous receptor

The Cells
The GeneBLAzer® PPAR alpha-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human peroxisome proliferator-activated receptor alpha (PPAR alpha) fused to the DNA-binding domain of GAL4 transiently transduced (via BacMam virus) into the GeneBLAzer® UAS-bla HEK293T cell line. GeneBLAzer® UAS-bla HEK 293T cells stably express a β-lactamase reporter gene under the transcriptional control of an upstream activator sequence (UAS).

The Assay
When an agonist binds to the LBD of the GAL4 (DBD)-PPAR alpha (LBD) fusion protein, the protein binds to the UAS, resulting in expression of β-lactamase. The FRET ratiometric readout from the live-cell substrate reduces the absolute and relative errors that can mask the underlying biological response of interest. PPAR alpha UAS-bla HEK 293T cells have been tested for assay performance using variable assay conditions, including cell number, stimulation time, substrate loading time and have been validated for Z′ and EC50 concentrations of GW7647. Additional testing data using alternate stimuli are also available. See the "validation packet" in the documents below to view the data.

Speed and Convenience
Because PPAR alpha UAS-bla HEK 293T cells are transiently transduced with BacMam virus, they are ready to be used in an assay without any additional cell culture steps. Just plate the cells and assay as outlined in the protocol.

Please email our Custom Services team to inquire about obtaining these cells at economical bulk scales.

Tango™ MCHR2-bla U2OS Cells

The Tango™ MCHR2-bla U2OS cells contain the human Melanin-concentrating hormone receptor 2 (MCHR2) 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™ MCHR2-bla U2OS cells are functionally validated for Z' and EC50 concentrations of I-TAC (CXCL11). In addition, Tango™ MCHR2-bla U2OS cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

For Academic pricing please login or contact us at discoverysciences@invitrogen.com.

GeneBLAzer™ ADRB1-CRE-bla CHO-K1 Cells

The GeneBLAzer ® ADRB1-CRE-bla CHO-K1 cells contain the human Adrenergic Beta-1 Receptor (ADRB1), (Accession # NM_000684) 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 ADRB1-CRE-bla CHO-K1 cells are functionally validated for Z'-factor and EC50 concentrations of (-) Denopamine. In addition, ADRB1-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™ M4-Gqo5-NFAT-bla CHO-K1 Cells

GeneBLAzer® M4 Gqo5-NFAT-bla CHO-K1 cells contain the human Acetylcholine (muscarinic) subtype 4 receptor (M4) stably integrated into the GeneBLAzer® Gqo5-NFAT-bla CHO-K1 cell line. GeneBLAzer® Gqo5-NFAT-bla CHO-K1 cells (Cat. no. K1536) contain a beta-lactamase reporter gene under control of the Nuclear Factor of Activated T-cells (NFAT) response element and the Chimeric G protein, Gqo5.

The GeneBLAzer® M4 Gqo5-NFAT-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of Cabachol. In addition, GeneBLAzer® M4 Gqo5-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 clinical implications of the M4 receptor are unknown, however studies with knockout mice suggest that M4 may have implications in dopaminergic function, karatinocyte migration and wound healing, anxiolysis, and analgesia. In addition, M4 may be important in memory circuits and have implications in Alzheimer's Disease. Additional information on the muscarinic receptors can be found in literature.

For Academic pricing please login or contact us at discoverysciences@invitrogen.com.

GeneBLAzer™ ADORA2A-CRE-bla CHO-K1 Cells

GeneBLAzer® ADORA2A-CRE-bla CHO-K1 cells contain the human Adenosine Receptor A2A (ADORA2A) 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® ADORA2A-CRE-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of NECA. In addition, GeneBLAzer® ADORA2A-CRE-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

ADORA2A has been shown to have many roles throughout the body. ADORA2A is involved in pain and inflammation regulation, inhibition of platelet aggregation, blood pressure regulation and contributes to ischemic brain damage. Selective antagonists of the ADORA2A pathway are being tested as adjuvants to dopaminergic drugs to treat Parkinson's disease and schizophrenia.

For Academic pricing please login or contact us at discoverysciences@invitrogen.com.

LanthaScreen™ STAT3 GripTite™ Cell Line

The JAK/STAT3 signaling pathway is known to be activated by cytokines such as IL-6. In this pathway, binding of IL-6 to its cell-surface receptors results in the activation of JAKs, which in turn phosphoactivate STAT3 proteins at a specific tyrosine residue (Y705). LanthaScreen® STAT3 GripTite™ is a human cell line that constitutively expresses a GFP-STAT3 fusion protein. The JAK/STAT signaling pathway is known to be functionally intact in this cell line; therefore, the GFP-STAT3 fusion protein serves as a substrate for IL-6-inducible phosphorylation. Using this cell line, a lytic immunoassay has been developed in which the phosphorylation state of GFP-STAT3 is detected in cell lysates using a terbium-labeled anti-pY705-STAT3 antibody in a time-resolved FRET (TR-FRET) readout.

The GFP-STAT3 DNA expression construct was transfected into the GripTite™ HEK 293 cell line using Lipofectamine™ 2000 transfection reagent, and the transfected cells were selected with blasticidin. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as a sorting marker. The assay utilizing this cell line is validated for EC50 and Z’-factors under optimized conditions using IL-6 as a ligand for JAK-mediated GFP-STAT3 phosphorylation. This assay has also been tested under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance, and assay equilibration time. An alternate "mix-and-read" assay format is also described.

Tango™ CHRM2-bla U2OS Cells

The Tango™ CHRM2-bla U2OS cells contain the human Cholinergic muscarinic 2 receptor (CHRM2) 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™ CHRM2-bla U2OS cells have been functionally validated for Z' factor and EC50 concentrations of Carbachol. In addition, Tango™ CHRM2-bla U2OS 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™ PAC1-CRE-bla CHO-K1 Cells

The GeneBLAzer® PAC1-CRE-bla CHO-K1 cells contain the human Adenylate Cyclase Activating Polypeptide 1 Receptor 1 (ADCYAP1R⁄PAC1) 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 PAC1-CRE-bla CHO-K1 cells are functionally validated for Z'-factor and EC50 concentrations of established ligands. In addition, PAC1-CRE-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary

BacMam Histone H3K27me3 Cellular Assay Kit (Invitrogen™)

The BacMam Histone H3K27me3 Cellular Assay is a high-throughput screening (HTS) compatible TR-FRET assay for the interrogation of trimethylation at lysine 27 on histone H3 in a cellular format. Baculovirus-mediated gene delivery (BacMam) provides a convenient tool for the expression of GFP-Histone H3 fusion protein in your cell background of interest. Coupled with a terbium (Tb)-labeled site-specific antibody, the resulting cellular assay can be used to identify inhibitors of methyltransferases and demethylases acting on lysine 27 of histone H3.

The kit includes:

• BacMam Histone H3 Reagent
• LanthaScreen® Tb-anti-histone H3K27me3 Antibody
• 6X LanthaScreen® Cellular Lysis Buffer
• Instrument Control Terbium TR-FRET kit

With the BacMam Histone H3K27me3 Cellular Assay Kit You Can:
• Use your cell background of choice with the portability of BacMam
• Identify more relevant inhibitors since the methyl transferases and demethylases are in their natural protein complexes
• Conserve precious cell samples with the miniaturizable homogenous assay format
• Improve data quality with the advantages of TR-FRET

Get More Physiologically Relevant Results
The BacMam Histone H3K27me3 Cellular Assay allows the investigation of trimethylation at lysine 27 on histone H3 with your choice of cellular backgrounds including primary cells. This enables screening for potential inhibitors of methyl transferases and demethylases associated with Lys27 trimethylation in their natural complexes in a physiologically relevant cell type.

BacMam and LanthaScreen® Convenience Saves Sample and Time
Assays are run in a fully homogenous, addition-only format without any of the washing, lysate transfer, or separation procedures required for traditional methods such as Western blotting and ELISA, making them ideal for HTS applications. By using the same BacMam reagent for GFP-Histone H3, other histone H3 modifications (Ph, Me, Ac) can be monitored just be exchanging the Terbium (Tb) labeled antibody. In addition, the application of the LanthaScreen® technology includes all the advantages of TR-FRET detection including reduced data noise, less interference from fluorescent compounds, and high sensitivity, allowing the use of fewer cells than traditional Western or ELISA methods.

GeneBLAzer™ AVPR1A CHO-K1 DA Assay Kit

This cell line is available as dividing cells by requesting a quote for K2186 from dividingcells@invitrogen.com

GeneBLAzer® AVPR1A CHO-K1 DA(Division Arrested) cells and AVPR1A-NFAT-bla CHO-K1 cells contain the human Arginine Vasopressin 1a (AVPR1A) receptor stably integrated into the CellSensor® NFAT-bla CHO-K1 cell line. CellSensor® NFAT-bla CHO-K1 cells (Cat. no.K1129) contain a beta-lactamase reporter gene under control of the nuclear factor of activated T-cells (NFAT) response element. 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 AVPR1A CHO-K1 DA cells and AVPR1A-NFAT-bla CHO-K1 cells are functionally validated for Z’-factor and EC50 concentrations of vasopressin, (Figure 1). In addition, AVPR1A-NFAT-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.

The family of receptors that recognize vasopressin and oxytocin consists of four members: vasopressin receptors 1a (AVPR1a), 1b (AVPR1b), and 2 (AVPR2), and the oxytocin (OT) receptor. AVPR1a, AVPR1b, and the OT receptor couple to the Gq/G11 class of G proteins and trigger the release of intracellular calcium when stimulated. AVPR2 receptors couple to Gs proteins and activate adenyl cyclase. All three AVPR receptors have high affinity for their endogenous ligand, vasopressin, but AVPR1b has less affinity for oxytocin than does AVPR1a or AVPR2. The hormones vasopressin and oxytocin are synthesized in the hypothalamus. Oxytocin causes contractions in the uterus during childbirth. The main functions of arginine vasopressin (AVP) are reabsorption of water by the kidneys and contraction of smooth muscle in arteries. This system may also play a role in memory and learning.


Storage:

GeneBLAzer® AVPR1A CHO-K1 DA Cells and AVPR1A-NFAT-bla CHO-K1 Cells are shipped on dry ice. Store in liquid nitrogen immediately upon receipt or thaw for immediate use.


Contents:
GeneBLAzer® AVPR1A 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:
•AVPR1A CHO-K1 DA cells (K1323A)
•LiveBLAzer™-FRET B/G Loading Kit, 70µg
•Solution D, 1ml
•2 protocols
•Certificate of Analysis
GeneBLAzer® AVPR1A 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:
•AVPR1A CHO-K1 DA cells (K1324A)
•1 protocol
•Certificate of Analysis

GeneBLAzer® AVPR1A-NFAT-bla CHO-K1 cells
(LiveBLAzer™-FRET B/G Loading Kit, Solution D and other materials are required separately; please refer to the protocol).
Includes:
•AVPR1A-NFAT-bla CHO-K1 cells (K1286)
•1 protocol
•Certificate of Analysis

GeneBLAzer™ T-REx™-G2A-NFAT-bla FreeStyle™ 293F Cells

GeneBLAzer® T-REx™ G2A-NFAT-bla Freestyle HEK 293F cells contain the human GPR132 (G2A), (Accession # NP_037477) stably integrated into the CellSensor® NFAT-bla Freestyle HEK 293F cell line. CellSensor® NFAT-bla Freestyle HEK 293F cells (Cat. no. K1725) contain a beta-lactamase reporter gene under control of the NFAT Response Element.

The GeneBLAzer® T-REx™ G2A-NFAT-bla Freestyle HEK 293F cells are functionally validated for Z' and EC50 concentrations of dDAVP. In addition, GeneBLAzer® GeneBLAzer® T-REx™ G2A-NFAT-bla Freestyle HEK 293F cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

G2A is an orphan GPCR. Upon overexpession G2A is constitutively active. This is an assay for the orphan GPCR based upon constitutive activity. To use the assay to detect inverse agonists, high amounts of doxycycline should be added. To use the assay to detect agonists, low amounts of doxycycline should be added.

For Academic pricing please login or contact us at discoverysciences@invitrogen.com.

GeneBLAzer™ GPR54 CHO-K1 DA Assay Kit

This cell line is available as dividing cells by requesting a quote for K1181 from dividingcells@invitrogen.com

GeneBLAzer® GPR54 CHO-K1 DA (Division Arrested) cells and GPR54-NFAT-bla CHO-K1 cells contain the human KiSS1 receptor (GPR54) stably integrated into the CellSensor® NFAT-bla CHO-K1 cell line. CellSensor® NFAT-bla CHO-K1 cells (Cat. no.K1078) contain a beta-lactamase reporter gene under control of the Nuclear Factor of Activated T-cells (NFAT) response element. 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 GPR54 CHO-K1 DA cells and GPR54-NFAT-bla CHO-K1 cells are functionally validated for Z’-factor and EC50 concentrations of Metastin, (Figure 1). In addition, GPR54-NFAT-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.

The GPR54 receptor has been shown to be involved in tumor progression and metastasis, and in the development and functional regulation of the gonads.


Storage:

GeneBLAzer® GPR54 CHO-K1 DA Cells and GPR54-NFAT-bla CHO-K1 Cells are shipped on dry ice. Store in liquid nitrogen immediately upon receipt or thaw for immediate use.


Contents:
GeneBLAzer® GPR54 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:
•GPR54 CHO-K1 DA cells
•LiveBLAzer™-FRET B/G Loading Kit, 70µg
•Solution D, 1ml
•2 protocols
•Certificate of Analysis
GeneBLAzer® GPR54 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:
•GPR54 CHO-K1 DA cells
•1 protocol
•Certificate of Analysis
GeneBLAzer® GPR54-NFAT-bla CHO-K1 cells
(LiveBLAzer™-FRET B/G Loading Kit, Solution D and other materials are required separately; please refer to the protocol).
Includes:
•GPR54-NFAT-bla CHO-K1 cells
•1 protocol
•Certificate of Analysis

GeneBLAzer™ P2RY4-NFAT-bla CHO-K1 Cells

GeneBLAzer® P2RY4-NFAT-bla CHO-K1 cells contain the human Purinergic Receptor P2Y4 (P2RY4), stably integrated into the CellSensor® NFAT-bla CHO-K1 cell line. CellSensor® NFAT-bla CHO-K1 cells (Cat.no.K1078) contain a beta-lactamase reporter gene under control of the NFAT Response Element. The GeneBLAzer® P2RY4-NFAT-bla CHO-K1 cells are functionally validated for Z’-factor and EC50 concentrations of UTP. In addition, GeneBLAzer® P2RY4-NFAT-bla CHO-K1 cells have been tested for assay performance under variable conditions.

Tango™ DRD1-bla U2OS Cells

The Tango™ DRD1-bla U2OS cells contain the human Dopamine Receptor 1 (DRD1) 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™ DRD1-bla U2OS cell line has been functionally validated for Z'-factor and EC50 concentrations of dihydrexidine. The DRD1 cells have been tested for assay performance under variable assay conditions. These data are available 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™ EDG3-Gα15-NFAT-bla HEK 293T Cells

GeneBLAzer® EDG3-Gα15-NFAT-bla HEK293T cells contain the human Endothelial Differentiation Receptor 3 (EDG3) stably integrated into the GeneBLAzer® EDG3-Gα15-NFAT-bla HEK293T cell line. GeneBLAzer® Gα15-NFAT-bla HEK293T cells (Cat. no. K1539) 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® EDG3-Gα15-NFAT-bla HEK293T cells are functionally validated for Z' and EC50 concentrations of S1P. In addition, GeneBLAzer® EDG3-Gα15-NFAT-bla HEK293T cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

EDG-3/S1P-3(Sphingosine-1-Phosphate-3) is a Gq/Gi/Go/Gα13/Gα12 coupled GPCR. EDG-3 has been shown to be responsible for bradycardia that has been induced in clinical trials as a side effect of the immunosuppressive drug FTY720 that targets EDG-1. EDG-3 has also been shown to induce vasodilation in response to sphingosylphosphorylcholine (SPC), sphingosine-1-phosphate (S1P), and lysosulfatide (LSF). The EDG-3 Receptor has also been shown to play a cooperative or redundant role in angiogenesis with EDG-1.

For Academic pricing please login or contact us at discoverysciences@invitrogen.com.

GeneBLAzer™ C5AR1-Gα15 CHO-K1 DA Assay Kit

The GeneBLAzer® C5AR1-Gα15 CHO-K1 DA (Division Arrested) cells contain the human Complement Component 5a Receptor 1 (C5AR1) (Accession # NM_000710) stably integrated into the GeneBLAzer® Gα15-NFAT-bla CHO-K1 cell line. GeneBLAzer® Gα15-NFAT-bla CHO-K1 (Cat # K1213) contains a beta-lactamase (bla) reporter gene under control of a NFAT response element and a promiscuous G Protein, Gα15, stably integrated into CHO-K1 cells.

The C5AR1-Gα15 CHO-K1 DA (Division Arrested) cells cells are functionally validated for Z'-factor and EC50 concentrations using C5a. In addition, C5AR1-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 in the Manuals and Brochures section. 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.

Tango™ EDG6-bla U2OS DA Assay Kit

The Tango™ EDG6-bla U2OS cells contain the human Endothelial Differentiation, Lysophosphatidic Acid G-protein-coupled Receptor, 6 (EDG6) 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™ EDG6-bla U2OS cells cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ EDG6-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. Division Arrested cells are irreversibly division arrested using a low-dose treatment of Mitomycin C, and have no apparent toxicity or change in cellular signal transduction.

GeneBLAzer™ CRHR2-CRE-bla CHO-K1 Cells

GeneBLAzer® CRHR2-CRE-bla CHO-K1 cells contain the human Corticotropin Releasing Factor Receptor 2 (CRHR2) 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® CRHR2-CRE-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of CRF. In addition, GeneBLAzer® CRHR2-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.

For Academic pricing please login or contact us at discoverysciences@invitrogen.com.

GeneBLAzer™ PTGER2-NFAT-bla CHO-K1 Cells

For academic pricing please login or contact us at discoverysciences@invitrogen.com.

GeneBLAzer® PTGER2-NFAT-bla CHO-K1 cells contain the human Prostaglandin E Receptor 2 (PTGER2), (Accession # NM_000956.2) stably integrated into the CellSensor® NFAT-bla CHO-K1 cell line. CellSensor® NFAT-bla CHO-K1 cells (Cat. no. K1534) contain a beta-lactamase reporter gene under control of the NFAT Response Element.

GeneBLAzer® PTGER2-NFAT-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

CellSensor™ AP-1-bla HEK293T Cell Line

The CellSensor® AP-1-bla HEK 293T Cell Line contains a beta-lactamase reporter gene under control of the AP-1 response element stably integrated into HEK 293T cells. The cell line was created through sorting by FACS of cells responsive to stimulation of the AP-1 pathway with phorbol 12-myristate 13-acetate (PMA). The cell line was validated for DMSO tolerance, incubation time with stimulant, and substrate loading conditions. The AP-1-bla HEK 293T cell line responds to agonist treatment as expected from literature and can be adapted for high throughput screening for agonists or antagonists of the AP-1 pathway with compound libraries. Candidate drugs can also be tested for dose response against this cell line. Academic and non-profit customers, please inquire for special pricing.

Tango™ ADORA1-bla U2OS DA Assay Kit

The Tango™ ADORA1-bla U2OS DA (Division Arrested) cells contain the human Adenosine A1 receptor (ADORA1) 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™ ADORA1-bla U2OS DA cells have been functionally validated for Z' factor and EC50 concentrations of NECA. In addition, Tango™ ADORA1-bla U2OS 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. 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.