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Tango™ GPR119-bla U2OS Cells

The Tango™ GPR119-bla U2OS cells contain the human G-protein Coupled Receptor 119 (GPR119) 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™ GPR119-bla U2OS cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ GPR119-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™ 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.

Tango™ F2RL1-bla U2OS Cells

The Tango™ F2RL1-bla U2OS cells contain the human Coagulation factor II (thrombin) receptor-like 1 (F2RL1) 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™ F2RL1-bla U2OS cells are functionally validated for Z' and EC50 concentrations of SLIGRL. In addition, Tango™ F2RL1-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™ 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.

GeneBLAzer™ ER Beta DA Assay Kit

GeneBLAzer®ER beta DA(Division Arrested) cells and ER beta-UAS-bla GripTite™ 293 cells contain the ligand-binding domain (LBD) of the human Estrogen receptor beta (ER beta) fused to the DNA-binding domain of GAL4 stably integrated in the GeneBLAzer®UAS-bla GripTite™ 293 cell line. GeneBLAzer®UAS-bla GripTite™ 293 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)-ER 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 ER beta DA cells and ER beta-UAS-bla GripTite™ 293 cells are functionally validated for Z' and EC50 concentrations of 17-beta Estradiol. In addition, ER beta-UAS-bla GripTite™ 293 cells have been tested for assay performance under variable conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time. Additional testing data using alternate stimuli are also available.

CellSensor™ c-Fos-bla HEK293T Cell Line

The c-fos-bla HEK 293T cell line contains a beta-lactamase reporter gene under control of the c-fos promoter stably integrated into HEK 293T cells. This cell line can be used to detect agonists/antagonists of the c-fos pathway. The c-fos-bla HEK 293T cells have been shown to be responsive to phorbol 12-myristate 13-acetate (PMA) stimulation. Academic and non-profit customers, please inquire for special pricing.

CellSensor™ LEF/TCF-bla HCT-116 Cell Line

Wnt signaling via Beta-catenin plays a central role in development and homeostasis. This pathway is invariably disrupted in colorectal tumors and commonly affected by mutation in other cancers. Wnt ligand binding and activating the Frizzled transmembrane receptors transduced the signal to a cytoplasmic protein, known as disheveled protein, which then inhibits the serine/threonine kinase Glycogen Synthase-3 Beta (GSK-3B). This signal leads to functional inactivation and dissociation of a multi-protein Beta -catenin destruction comples, which is made up of the tumor suppressor protein Adenomatous Polyposis Coli (APC), GSK-3B and a scaffold of Axin. This results in dephosphorylation and dissociation of Beta -catenin. The unphosphorylated b-catenin is stabilized and accumulates in the cytoplasm of the cell. Beta -catenin then associates with the T-Cell Factor (TCF)/Lymphoid Enhancer Factor (LEF) family of transcription factors in the nucleus leading to transcription and expression of target genes, such as c-Myc, c-jun, Fra and cyclin D1. The CellSensor™ LEF/TCF-bla HCT-116 cell line contains a beta-lactamase reporter gene under control of the LEF/TCF stably integrated into HCT-116 cells. HCT-116 is a colon cancer cell line carrying a gain-of-function mutation in beta-catenin gene (deletion of amino acid Serine45), which prevents beta-catenin protein degradation leading to constitutive activation of downstream genes. Thus, this cell line constitutively expresses beta-lactamase, which may be further stimulated or inhibited, e.g., using Stealth RNAi™ siRNA. This cell line has also been tested under various experimental conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time. Academic and non-profit customers, please inquire for special pricing.

GeneBLAzer™ EDG3-Gα15 HEK 293T DA Assay Kit

This cell line is available as dividing cells by requesting a quote for K1234 from dividingcells@invitrogen.com

GeneBLAzer® EDG3-Gα15 HEK 293T DA (Division Arrested) cells and EDG3-Gα15-NFAT-bla HEK 293T cells contain the human EDG3 receptor stably integrated in the GeneBLAzer® Gα15-NFAT-bla HEK 293T cell line. GeneBLAzer® Gα15-NFAT-bla HEK 293T cells (#K1212) contain a beta-lactamase reporter gene under control of the Nuclear Factor of Activated T-cells (NFAT) response element, and stably express the promiscuous G-protein, Gα15. 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 EDG3-Gα15 HEK 293T DA cells and EDG3-Gα15-NFAT-bla HEK 293T cells are functionally validated for Z’-factor and EC50 concentrations of Sphingosine-1-phosphate (S1P), (Figure 1). In addition, EDG3-Gα15-NFAT-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.

EDG-3(Endothelial-differentiation-gene-3)/S1P-3(Sphingosine-1-Phosphate-3) is a Gq/Gi/Go/G13/G12 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

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™ 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.

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.

GeneBLAzer™ PPAR gamma DA Assay Kit

GeneBLAzer®PPAR gamma DA (Division Arrested) cells and PPAR gamma-UAS-bla HEK 293H cells contain the ligand-binding domain (LBD) of the human Peroxisome Proliferator-Activated Receptor-gamma (PPAR gamma) fused to the DNA-binding domain of GAL4 stably integrated in the GeneBLAzer®UAS-bla HEK 293H cell line. GeneBLAzer®UAS-bla HEK 293H 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)-PPAR gamma (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 PPAR gamma DA cells and PPAR gamma-UAS-bla HEK 293H cells are functionally validated for Z' and EC50 concentrations of Rosiglitazone. In addition, PPAR gamma-UAS-bla HEK 293H cells have been tested for assay performance under variable conditions, including DMSO concentration, cell number, and stimulation time.

Tango™ SSTR2-bla U2OS Cells

The Tango™ SSTR2-bla U2OS cells contain the human Somatostatin Receptor 2 (SSTR2) 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™ SSTR2-bla U2OS cells have been functionally validated for Z' factor and EC50 concentrations of Somatostatin-14. In addition, Tango™ SSTR2-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.

Tango™ NPY1R-bla U2OS Cells

The Tango™ NPY1R-bla U2OS cells contain the human Neuropeptide Y Receptor Y1 (NPY1R) 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™ NPY1R-bla U2OS cells are functionally validated for Z' and EC50 concentrations of Neuropeptide-Y. In addition, Tango™ NPY1R-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™ OPRD1-bla U2OS Cells

The Tango™ OPRD1 -bla U2OS cells contain the human Opioid Receptor Delta 1 (OPRD1) 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™ OPRD1-bla U2OS cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ OPRD1-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.