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GeneBLAzer™ ADRA1A-NFAT-bla CHO-K1 Cells

The GeneBLAzer® ADRA1A-NFAT-bla CHO-K1 cells contain the human Adrenergic Alpha-1A Receptor (ADRA1A), (Accession # NM_000680) 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.

Both ADRA1A-NFAT-bla CHO-K1 cells are functionally validated for Z'-factor and EC50 concentrations of Phenylephrine. In addition, ADRA1A-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™ HCRTR1-NFAT-bla CHO-K1 Cells

The GeneBLAzer® HCRTR1-NFAT-bla CHO-K1 cells contain the human Hypocretin (Orexin) Receptor 1 (HCRTR1), (Accession # NM_001525) 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 HCRTR1-NFAT-bla CHO-K1 cells are functionally validated for Z'-factor and EC50 concentrations of Orexin A. In addition, HCRTR1-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.

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.

GeneBLAzer™ ADRA1B-NFAT-bla CHO-K1 Cells

The GeneBLAzer® ADRA1B-NFAT-bla CHO-K1 cells contain the human Complement Component 5a Receptor 1 (ADRA1B) 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 ADRA1B-NFAT-bla CHO-K1 cells are functionally validated for Z'-factor and EC50 concentrations using established ligands. In addition, ADRA1B-NFAT-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

GeneBLAzer™ RARβ-UAS-bla HEK 293T Cells

RAR beta-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human retinoic acid receptor beta (RAR beta) 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)-RAR beta (LBD) fusion protein, the protein binds to the UAS, resulting in expression of beta-lactamase. RAR beta-UAS-bla HEK 293T cells are functionally validated for Z' and EC50 concentrations of all-trans retinoic acid (ATRA). In addition, RAR beta-UAS-bla HEK 293T cells have been tested for assay performance under variable conditions, including DMSO concentration, cell number, and stimulation time.

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.

CellSensor™ NFAT CHO-K1 DA Assay Kit

The CellSensor® NFAT CHO-K1 DA (Division Arrested) cells contain a beta-lactamase (bla) reporter gene under control of a Nuclear Factor of Activated T Cells (NFAT) response element stably integrated into CHO-K1 cells. Fluorescence activated cell sorting (FACS) was used to isolate clones responsive to stimulation of the NFAT pathway by PMA (Phorbol 12-Myristate 13-Acetate) and Thapsagargin (Thaps). The cell line was validated for DMSO tolerance, cell number, stimulation time, substrate loading conditions, and EC50 concentration of Thaps at a fixed concentration of PMA. 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™ 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.

GeneBLAzer™ ADRB2-CRE-bla CHO-K1 Cells

The GeneBLAzer® ADRB2-CRE-bla CHO-K1 cells contain the human Adrenergic Beta-2 Receptor (ADRB2), (Accession # NM_000024.3) stably integrated into the CellSensor® CRE-bla CHO-K1 cell line. CellSensor® CRE-bla CHO-K1 cells (Cat # K1129) contain a beta-lactamase (bla) reporter gene under control of the cAMP response element (CRE).

The ADRB2-CRE-bla CHO-K1 cells are functionally validated for Z'-factor and EC50 concentrations of Isoproterenol. In addition, ADRB2-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™ MR-UAS-bla HEK 293T Cells

MR-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human Mineralcorticoid Receptor (MR) fused to the NA-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)-MR (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. MR-UAS-bla HEK 293T cells are functionally validated for Z' and EC50 concentrations of aldosterone. In addition, MR-UAS-bla HEK 293T cells have been tested for assay performance under variable conditions, including DMSO concentration, cell number, and substrate loading time. Additional testing data using alternate stimuli are also available.

Tango™ CNR2-bla U2OS DA Assay Kit

The Tango™ CNR2-bla U2OS DA (Division Arrested) cells and Tango™ CNR2-bla U2OS cells contain the human Cannabinoid Receptor 2 (CNR2) 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™ CNR2-bla U2OS cells and the Tango™ CNR2-bla U2OS DA cells have been functionally validated for Z' factor and EC50 concentrations of CP-55940. In addition, Tango™ CNR2-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™ OPRL1-bla U2OS Cells

The Tango™ OPRL1-bla U2OS cells contain the human Opioid Receptor-Like 1 (OPRL1) 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™ OPRL1-bla U2OS cells have been functionally validated for Z’ factor and EC50 concentrations of established ligands. In addition, Tango™ OPRL1-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™ NFKB-bla Freestyle™ HEK293F Cell Line

Nuclear Factor kappaB (NFκB) signaling regulates genes involved in apoptosis, viral defense, cancer, inflammation, and autoimmune disease. The CellSensor® NFκB-bla FreeStyle™ 293F Cell Line is responsive to tumor necrosis factor alpha (TNFα) and can be used to probe the NFκB signaling pathway. This cell line was validated for DMSO tolerance, incubation time with stimulant and substrate loading conditions. The CellSensor® NFκB-bla FreeStyle™ 293F Cell Line contains a beta-lactamase reporter gene under the control of NFκB response element. The construct was transduced into FreeStyle™ 293F cells by lentivirus. This cell line is a pool isolated for response to TNFα by flow cytometry. It responds to agonist treatment as expected from literature. Academic and non-profit customers, please inquire for special pricing.

GeneBLAzer™ PROKR2-Gqi5-NFAT-bla CHO-K1 Cells

GeneBLAzer® PROKR2-Gqi5-NFAT-bla CHO-K1 cells contain the human prokineticin receptor 2 (PROKR2), (Accession # NM_144773) stably integrated into the GeneBLAzer® Gqi5-NFAT-bla CHO-K1 cell line. GeneBLAzer® Gqi5-NFAT-bla CHO-K1 cells (Cat No K1725) contain a beta-lactamase reporter gene under control of the NFAT Response Element. The GeneBLAzer® PROKR2-Gqi5-NFAT-bla CHO-K1 cells are functionally validated for Z’-factor and EC50 concentrations of Prokineticin (EG-VGEF)

GeneBLAzer™ TRβ-UAS-bla HEK 293T Cells

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. 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.