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Tango™ GPR119-bla U2OS DA Assay Kit

The Tango™ GPR119-bla U2OS DA 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 DA cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ GPR119-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 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™ NTSR1 CHO-K1 DA Assay Kit

The GeneBLAzer® NTSR1 CHO-K1 DA cells contain the human Neurotensin Receptor 1 (NTSR1) stably integrated into the GeneBLAzer® Gqo5-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® NTSR1 CHO-K1 DA cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, GeneBLAzer® NTSR1 CHO-K1 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.

T-REx™ Tango™ P2RY14-bla U2OS Cells

T-REx™ Tango™ P2RY14-bla U2OS cells contain the human purinergic receptor P2Y14 (P2RY14) under doxycycline inducible control 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 T-REx™ Tango™ P2RY14-bla U2OS cells have been functionally validated for Z’ factor and EC50 concentrations of UDP-glucose. In addition, T-REx™ Tango™ P2RY14-bla U2OS cells have been tested for assay performance under variable conditions.

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.

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.

Tango™ GPR44 (CRTH2)-bla U2OS Cells

Tango™ GPR44-bla U2OS cells contain the human G protein coupled receptor 44 (GPR44) 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™ GPR44-bla U2OS cells have been functionally validated for Z’ factor and EC50 concentrations of Indomethacin. In addition, Tango™ GPR44-bla U2OS cells have been tested for assay performance under variable conditions.

GeneBLAzer™ LTB4R-Gα15-NFAT-bla CHO-K1 Cells

GeneBLAzer® LTB4R-Gα15-NFAT-bla CHO-K1 cells contain the human Leukotriene B4 Receptor (LTB4R), (Accession # NM_181657) 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 (bla) reporter gene under control of the NFAT Response Element.

GeneBLAzer® LTB4R-Gα15-NFAT-bla CHO-K1 cells are functionally validated for Z'-factor and EC50 concentrations of LTB4. In addition, GeneBLAzer® LTB4R-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.

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.

LanthaScreen™ ERK2 A375 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). A number of constitutive active kinase mutants within the MAP kinase pathway have been implicated in oncogenesis. One of these mutants is BRAF(V600E), which is the most predominant oncogenic BRAF mutant. The human melanoma cell line A375 endogenously expresses BRAF(V600E), which leads to the constitutive activation of the MAP kinase pathway and phosphorylation of Erk2 in the absence of ligands. LanthaScreenTM Erk2 A375 constitutively expresses GFP-Erk2 under control of a CMV promotor. 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. This cell line can be used to evaluate compound activity against BRAF(V600E). GFP-Erk2 lentivirus was transduced into A375 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.

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.

CellSensor™ NFAT-bla Jurkat Cell Line

The CellSensor® NFAT-bla Jurkat cell line contains a beta-lactamase reporter gene under control of a Nuclear Factor of Activated T Cells (NFAT) response element stably integrated into Jurkat cells. This cell line can be used as a parental cell line to build specific G-protein coupled receptor (GPCR) assays after transfection of additional genes of interest. NFAT-bla Jurkat cells have been shown to be responsive to phorbol 12-myristate 13-acetate (PMA) and Ionomycin stimulation. Additional testing data using alternate stimuli is available upon request. Academic and non-profit customers, please inquire for special pricing.

Tango™ M1-bla U2OS Cells

Tango™ M1-bla U2OS cells contain the human muscarinic M1 receptor (CHRM1) 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™ M1-bla U2OS cells have been functionally validated for Z’ factor and EC50 concentrations of oxotremorine M. In addition, Tango™ M1-bla U2OS cells have been tested for assay performance under variable conditions.

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™ CXCR4-bla U2OS Cells

The Tango™ CXCR4-bla U2OS cells contain the human Chemokine (C-X-C motif) receptor 4 (CXCR4) 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™ CXCR4-bla U2OS cells are functionally validated for Z' and EC50 concentrations of SDF1α. In addition, Tango™ CXCR4-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.