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GeneBLAzer™ VDR-UAS-bla HEK 293T Cells

VDR-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human Vitamin D receptor (VDR) fused to the DNAbinding 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)-VDR (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. VDR-UAS-bla HEK 293T cells are functionally validated for Z' and EC50 concentrations of Calcitriol (1α, 25-dihydroxyvitamin D3). In addition, VDR-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.

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™ CALCRL:RAMP3-CRE-bla FreeStyle™ 293F Cells

The GeneBLAzer® CALCRL:RAMP3-CRE-bla FreeStyle™ 293F cells contain the human Amylin Receptor 2 (CALCRL:RAMP3), (Accession # NM_005795.3 NM_005856.1) stably integrated into the CellSensor® CRE-bla Freestyle™ 293F cell line. CellSensor® CRE-bla Freestyle™ 293F cells (Cat # K1130) contain a beta-lactamase (bla) reporter gene under control of the cAMP response element (CRE).

CALCRL:RAMP3-CRE-bla FreeStyle™ 293F cells are functionally validated for Z'-factor and EC50 concentrations of Adrenomedullin(1-52). In addition, CALCRL:RAMP3-CRE-bla FreeStyle™ 293F 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.

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.

Tango™ OPRK1-bla U2OS Cells

The Tango™ OPRK1-bla U2OS cells contain the human Opiod Receptor Kappa 1 (OPRK1) 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™ OPRK1-bla U2OS cells cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ OPRK1-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™ CCR5-bla U2OS Cells

The Tango™ CCR5-bla U2OS cells contain the human Chemokine (C-C Motif) Receptor 5 (CCR5) 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™ CCR5-bla U2OS cells are functionally validated for Z' and EC50 concentrations of MIP-1a. In addition, Tango™ CCR5-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™ CRHR1 CHO-K1 DA Assay Kit

This cell line is available as dividing cells by requesting a quote for K1263 from dividingcells@invitrogen.com

GeneBLAzer® CRHR1 CHO-K1 DA (Division Arrested) cells and CRHR1-CRE-bla CHO-K1 cells contain the human Corticotropin Releasing Factor 1 (CRHR1) 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 CRHR1 CHO-K1 DA cells and CRHR1-CRE-bla CHO-K1 cells are functionally validated for Z’-factor and EC50 concentrations of corticotropin releasing factor (CRF), (Figure 1). In addition, CRHR1-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.

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 has revealed no major differences between them.

Storage:
GeneBLAzer® CRHR1 CHO-K1 DA Cells and CRHR1-CRE-bla CHO-K1 Cells are shipped on dry ice. Store in liquid nitrogen immediately upon receipt or thaw for immediate use.


Contents:
GeneBLAzer® CRHR1 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:
•CRHR1 CHO-K1 DA cells
•LiveBLAzer™-FRET B/G Loading Kit, 70µg
•Solution D, 1ml
•2 protocols
•Certificate of Analysis
GeneBLAzer® CRHR1 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:
•CRHR1 CHO-K1 DA cells
•1 protocol
•Certificate of Analysis
GeneBLAzer® CRHR1-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:
•CRHR1-CRE-bla CHO-K1 cells
•1 protocol
•Certificate of Analysis

GeneBLAzer™ P2RY6 CHO-K1 DA Assay Kit

This cell line is available as dividing cells by requesting a quote for K1253 from dividingcells@invitrogen.com

GeneBLAzer® P2RY6 CHO-K1 DA (Division Arrested) cells and P2RY6-NFAT-bla CHO-K1 cells contain the human purinergic receptor P2, G protein-coupled, 6 (P2RY6) receptor stably integrated into the CellSensor® NFAT-bla CHO-K1 cell line. CellSensor® NFAT-bla CHO-K1 cells (Cat. #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 P2RY6 CHO-K1 DA cells and P2RY6-NFAT-bla CHO-K1 cells are functionally validated for Z’-factor and EC50 concentrations of Uridine 5’-diphosphate (UDP); (Figure 1). In addition, P2RY6-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.

P2RY6 has been found in a variety of rat tissue such as; placenta, spleen, thymus, lung, stomach, intestine, mesentery, heart, and aorta. P2RY6 is activated by UDP, which functions as a full agonist. P2RY6 is activated either weakly or not at all by additional extracellular nucleotides such as UTP, ATP, and ADP. P2RY6 has been implicated to be involved in ion transport, vasodilatation, and contraction of smooth muscle.


Storage:

GeneBLAzer® P2RY6 CHO-K1 DA Cells and P2RY6-NFAT-bla CHO-K1 Cells are shipped on dry ice. Store in liquid nitrogen immediately upon receipt or thaw for immediate use.


Contents:
GeneBLAzer® P2RY6 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:
•P2RY6 CHO-K1 DA cells
•LiveBLAzer™-FRET B/G Loading Kit, 70µg
•Solution D, 1ml
•2 protocols
•Certificate of Analysis
GeneBLAzer® P2RY6 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:
•P2RY6 CHO-K1 DA cells
•1 protocol
•Certificate of Analysis
GeneBLAzer® P2RY6-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:
•P2RY6-NFAT-bla CHO-K1 cells
•1 protocol
•Certificate of Analysis

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.

CellSensor™ ISRE-bla RA-1 Cell Line

The CellSensor® ISRE-bla RA-1 Cell Line contains the beta-lactamase (bla) gene under the control of the interferon-stimulated response element (ISRE). This cell line was engineered by lentiviral transduction of an ISRE-bla construct into RA-1 cells. Flow cytometry was then used to isolate a pool of cells responsive to polyinosinic-polycytidylic acid (poly I:C) stimulation. ISRE-bla RA-1 cells have been tested for assay performance using variable assay conditions, including DMSO concentration, cell number, stimulation time, substrate loading time, as well as validated for Z'-factor and EC50 concentrations of poly I:C. Additional testing data using alternate stimuli are also available. Academic and non-profit customers, please inquire for special pricing.

GeneBLAzer™ LXR alpha DA Assay Kit

GeneBLAzer®LXR alpha DA (Division Arrested) cells and 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. 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 LXR alpha DA cells and 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™ GRPR CHO-K1 DA Assay Kit

The GeneBLAzer® GRPR-NFAT-bla CHO-K1 cells contain the human Gastrin Releasing Peptide Receptor (GRPR) 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 GRPR-NFAT-bla CHO-K1 cells are functionally validated for Z'-factor and EC50 concentrations using established ligands. In addition, GRPR-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™ RAR alpha DA Assay Kit

The GeneBLAzer®RAR alpha DA (Division Arrested) and RAR alpha-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human retinoic acid receptor alpha fused to the DNA-binding domain of GAL4 stably integrated in the GeneBLAzer®UAS-bla HEK293T cell line. GeneBLAzer®UAS-bla HEK 293T cells (catalog#K1104) stably express a beta-lactamase reporter gene under the transcriptional control of a 7x Upstream Activator Sequence (UAS). Transcription from the 7xUAS is activated by the binding of the GAL4 transcription factor DNA-binding-domain (DBD). The GAL4-DBD is expressed as a fusion protein with the ligand binding domain (LBD) of RAR alpha. When an agonist binds to the LBD of the GAL4(DBD)-RAR alpha(LBD) fusion protein it translocates to the nucleus where it binds to the 7x UAS inducing transcription of beta-lactamase. Division Arrested (DA) cells are available in an Assay Kit (which includes cells and sufficient substrate to analyze 1 x 384-well plate). 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 RAR alpha DA cells and RAR alpha-UAS-bla HEK 293T cells have been tested for assay performance using variable assay conditions, including DMSO concentration, cell number, stimulation time, substrate loading time and have been validated for Z' and EC50 concentrations of all-trans retinoic acid. Additional testing data using alternate stimuli are also available.

Tango™ CXCR7-bla U2OS Cells

The Tango™ CXCR7-bla U2OS cells contain the human Chemokine (C-X-C motif) receptor 7 (CXCR7) 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™ CXCR7-bla U2OS cells are functionally validated for Z' and EC50 concentrations of SDF1a. In addition, Tango™ CXCR7-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.

CellSensor™ irf1-bla CTLL-2 Cell Line

Jak/Stat signaling pathways play essential roles in the 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 CTLL2 Cell Line contains a beta - lactamase reporter gene under control of the irf - 1 response element stably integrated into CTLL2 cells. CTLL2 cells are a clone of cytotoxic T cells derived from a C57BL/6 mouse, and are cell - growth dependent on mouse Interleukin - 2 (mIL - 2). This cell line is validated for EC50 and Z' - factor under optimized conditions using mIL - 2. This cell line has also been tested under variable assay conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time. Responsiveness to pathway inhibitors was also tested. CellSensor® irf1 - bla CTLL2 cells were stimulated with mIL - 2 in triplicate over the indicated concentration range in a 384 - well format. Cells were incubated for 5 hrs with agonist 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. The 460/530 ratios for each replicate plotted against the indicated concentrations of mIL - 2. CellSensor® irf1 - bla CTLL2 cells were treated with Jak Inhibitor 1 over the indicated concentration range in a 384 - well format for 30 min. Cells were then incubated for 5 hrs with mIL - 2 agonist (1 ng/ml) in 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, converted to percent inhibition relative to a set of controls (unstimulated cells and EC80 stimulated cells), and plotted against the indicated concentrations of Jak Inhibitor 1. Academic and non-profit customers, please inquire for special pricing.