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CellSensor™ NFKB-bla HEK293T Cell Line

The CellSensor® NFκB-bla HEK 293T cell line contains a beta-lactamase reporter gene under the control of the Nuclear Factor kappa B (NFκB) response element. To obtain the cell line, the NFκB-bla construct was transduced into HEK 293T cells by lentivirus. Subsequent flow cytometry was used to isolate clones responsive to Tumor Necrosis Factor alpha (TNFα). The cell line has been validated for DMSO tolerance, stimulation time, substrate loading time, and EC50 concentration of TNF&alpha. This cell line is responsive to TNFα and can be used to probe NFκB signaling pathways, including those that are involved in apoptosis, viral defense, cancer, inflammation, and autoimmune disease. Academic and non-profit customers, please inquire for special pricing.

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.

GeneBLAzer™ M5-NFAT-bla CHO-K1 Cells

GeneBLAzer® M5-NFAT-bla CHO-K1 cells contain the human Acetylcholine (Muscarinic) Receptor 5 (M5) 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® M5-NFAT-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of Carbachol. In addition, GeneBLAzer® M5-NFAT-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

Muscarinic acetylcholine receptor subtype 5 (M5) has an unknown physiological relevance primarily because of low expression levels and the lack of M5 receptor-selective ligands. However, knockout studies have suggested that M5 is involved in modulation of central dopamine function and the tone of cerebral blood vessels.

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

CellSensor™ SIE-bla THP-1 Cell Line

The CellSensor® SIE-bla THP-1 cell line contains a beta-lactamase reporter gene under control of the Sis Inducible Element (SIE), stably integrated into THP-1 cells. To construct this cell line, the SIE-bla construct was transduced into THP-1 cells by lentivirus. Flow cytometry was used to isolate cells responsive to Interferon Gamma (IFNγ). 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® SIE-bla THP-1 cell line is responsive to IFNγ and can be used to analyze the JAK/STAT pathway using the GeneBLAzer® Loading Substrates. Academic and non-profit customers, please inquire for special pricing.

GeneBLAzer™ Gα15-NFAT CHO-K1 DA Assay Kit

The GeneBLAzer® Gα15-NFAT CHO-K1 DA (Division Arrested) cell line contains the promiscuous G protein, Gα15, stably integrated into the CellSensor® NFAT-bla CHO-K1 (Cat #K1078) cell line. NFAT-bla CHO-K1 cells contain a beta-lactamase reporter gene under control of an NFAT response element stably integrated into CHO-K1 cells. 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.

This product can serve as a negative control in screening assays performed with specific G-protein coupled receptor (GPCR) DA cells that were made from this same background. These cells can also be used in other assays where a sensitive readout to intracellular changes in calcium is needed.

CellSensor™ NFκB-bla RA 1 Cell Line

The CellSensor® NFκB-bla RA-1 cell line contains a beta-lactamase reporter gene under control of the NFκB response element stably integrated into Ramos 1 (RA-1) cells. To construct this cell line, the NFκB -bla construct was transduced into RA-1 cells by lentivirus. Flow cytometry was used to isolate cells responsive to Tumor Necrosis Factor (TNFThis cell line was validated for DMSO tolerance, cell number, stimulation time, substrate loading time, as well as Z´ and EC50 concentrations for TNF. The CellSensor® NFκB-bla RA-1 cell line is responsive to TNFand can be used to probe NFκB signaling pathways, which are involved in the regulation of apoptosis, viral replication, tumorigenesis, inflammation, and various autoimmune diseases. This cell line can be adapted for high-throughput screening for agonists or antagonists with compound libraries. Candidate drugs can be tested for dose response against this cell line. Academic and non-profit customers, please inquire for special pricing.

GeneBLAzer™ LXRα-UAS-bla HEK 293T Cells

LXR alpha-UAS-bla HEK 293T 293 cells contain the ligand-binding domain (LBD) of the human Liver-X receptor-alpha(LXR alpha) fused to the DNA-binding 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)-LXR alpha (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. LXR alpha-UAS-bla HEK 293T 293 cells are functionally validated for Z' and EC50 concentrations of TO901317. In addition, LXR alpha-UAS-bla HEK 293T 293 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™ AVPR2-CRE-bla CHO-K1 Cells

GeneBLAzer® AVPR2 CRE-bla CHO-K1 cells contain the human Arginine Vasopressin 2 (AVPR2) receptor 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® AVPR2 CRE-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of dDAVP. In addition, GeneBLAzer® AVPR2 CRE-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

Agonists and antagonists of the AVPR2 receptors are in various stages of clinical trials for the treatment of hypertension and as diuretics.

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

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.

Tango™ CNR1-bla U2OS Cells

The Tango™ CNR1-bla U2OS cells contain the human Cannabinoid Receptor 1 (CNR1) 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™ CNR1 cell line has been functionally validated for Z'-factor and EC50 concentrations of CP 55,940. The CNR1 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.

CellSensor™ HRE-bla HCT-116 Cell Line

Hypoxia is an almost universal hallmark of solid tumors. Adaptation to hypoxia is critical for tumor survival and growth, and is mediated largely by transcriptional activation of genes that facilitate short - (e.g., glucose transport) and long - term (e.g., angiogenesis) adaptive mechanisms. This coordinated homeostatic response is mediated in large part through the activation of the heterodimeric transcription factor hypoxia - inducible factor 1 (HIF - 1). Under conditions of normal oxygenation, the regulated HIF - 1α is hydroxylated and degraded by the proteasome system. As oxygen becomes rate limiting, hydroxylation diminishes and HIF - 1α accumulates and heterodimerizes with the constitutively present α - subunit. The binding of this complex to the cognate hypoxia - response element (HRE) results in transcriptional activation of genes containing such elements within promoter or enhancer elements. The CellSensor® HRE-bla HCT - 116 Cell Line contains a beta - lactamase reporter gene under control of the HRE stably integrated into HCT - 116 cells. This cell line is validated for EC50 and Z'; - factor under optimized conditions using deferoxamine (DFO) and Cobalt Chloride. This cell line has also been tested under various experimental conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time. CellSensor ® HRE-bla HCT - 116 cells were stimulated with deferoxamine (DFO) over the indicated concentration range in a 384 - well format. Cells were incubated for 24 hrs with agonist and 0.5% DMSO and then combined with LiveBLAzer™ - FRET B/G Substrate (CCF4 - AM) for 2 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 DFO. CellSensor® HRE - bla HCT - 116 cells were stimulated with Cobalt Chloride (CoCl2)over the indicated concentration range in a 384 - well format. Cells were incubated for 24 hrs with agonist and 0.5% DMSO and then combined with LiveBLAzer™ - FRET B/G Substrate (CCF4 - AM) for 2 hrs. Fluorescence emission values at 460 nm and 530 nm were obtained using a standard fluorescence plate reader. Response ratios were plotted against the indicated concentrations of CoCl2. Academic and non-profit customers, please inquire for special pricing.

LanthaScreen™ ERK2 U2OS Cell Line

The Map Kinase (MapK) signal transduction cascade is activated by growth factors such as EGF, PDGF, and HGF. Binding of these factors to their respective cell surface receptors results in the initiation of receptor tyrosine kinase activity, which leads to the sequential phospho-activation of downstream kinases such as Ras, Raf, MEK, and Erk1⁄2. Activated MEK phosphorylates Erk2 (Mapk1) proteins at a specific Thr-Tyr motif (Thr⁄Tyr 185⁄187). LanthaScreenTM Erk2 U2OS is a human cell line which constitutively expresses GFP-Erk2. The MapK signaling pathway is functionally intact in this cell line, therefore the GFP-Erk2 fusion protein serves as a substrate for the growth-factor-inducible phosphorylation by MEK. Using this cell line, a homogenous immuno-assay has been developed in which the phosphorylation state of GFP-Erk2 is detected in cell lysates using a terbium-labeled anti-pTpY-185⁄187-Erk2 antibody, in a time-resolved FRET (TR-FRET) readout. GFP-Erk2 lentivirus was transduced into U2OS cells, followed by selection with Blasticidin. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as sorting marker. Using the lytic TR-FRET immuno-assay, this cell line is validated for EC50 and Z’ under optimized conditions using EGF as a ligand for GFP-Erk2 phosphorylation. This assay has also been tested for assay performance under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance and cell lysis⁄equilibration time. Additional information using alternate stimuli and small molecule inhibitor is also provided.

GeneBLAzer™ P2RY11-NFAT-bla CHO-K1 Cells

GeneBLAzer® P2RY11-NFAT-bla CHO-K1 cells contain the human Purinergic receptor P2, G protein-coupled, 11 (P2RY11) 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® P2RY11-NFAT-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of ATP. In addition, GeneBLAzer® P2RY11-NFAT-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

P2RY11 expression is broadly distributed and has been found in the brain, pituitary, lymphocytes, spleen, intestines, macrophages, lung, stomach, adipose tissue, pancreas, kidney, prostate, heart, placenta, liver, and skeletal muscle. Differentiation of human promyelocytic HL60 cells and maturation of monocyte-derived dendritic cells have been shown to be mediated by ATP activation of P2RY11.

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

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™ Jurkat Control Kit

Th e GeneBLAzer® Jurkat Control Kit contains constitutively expressing (CMV-bla) and wildtype Jurkat cells to provide positive and negative controls, respectively, for evaluating beta-lactamase activity (Figure 1).
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