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LanthaScreen™ STAT1 U2OS Cell Line

The JAK/STAT1 signaling pathway is known to be activated by type I/II interferons such as interferon-gamma and interferon-alpha. In this pathway, binding of these cytokines to their respective cell-surface receptors results in the activation of JAKs, which in turn phosphoactivate STAT1 proteins at a specific tyrosine residue (Y701). LanthaScreen® STAT1 U2OS is a human cell line that constitutively expresses a GFPSTAT1 fusion protein. The JAK/STAT1 signaling pathway is known to be functionally intact in this cell line; therefore, the GFP-STAT1 fusion protein serves as a substrate for the IFN-gamma-inducible phosphorylation by JAKs. Using this cell line, a homogeneous immunoassay has been developed in which the phosphorylation state of GFP-STAT1 is detected in cell lysates using a terbium-labeled anti-pY701-STAT1 antibody in a time-resolved FRET (TR-FRET) readout. The GFP-STAT1 DNA expression construct was transfected into U2OS cells using Lipofectamine™ LTX Reagent, followed by selection with blasticidin. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as a sorting marker. Using the lytic TR-FRET immunoassay, this cell line is validated for EC50 and Zfi-factors under optimized conditions using IFN-gamma as a ligand for JAK-mediated GFP-STAT1 phosphorylation. This assay has also been tested for assay performance under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance, and assay development time. Additional information using alternate stimuli and alternate assay protocols is available.

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™ ADORA2A CHO-K1 DA Assay Kit

This cell line is available as dividing cells by requesting a quote for K1245 from dividingcells@invitrogen.com

GeneBLAzer® ADORA2A CHO-K1 DA (Division Arrested) cells and ADORA2A-CRE-bla CHO-K1 cells contain the human Adenosine A2a (ADORA2A) 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 ADORA2A CHO-K1 DA cells and ADORA2A-CRE-bla CHO-K1 cells are functionally validated for Z’-factor and EC50 concentrations of 5’-adenosine (NECA); (Figure 1). In addition, ADORA2A-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.

The Adenosine A2a Receptor (ADORA2A) has been shown to have many roles throughout the body. ADORA2A is involved in pain and inflammation regulation, inhibition of platelet aggregation, blood pressure regulation and contributes to ischemic brain damage. Selective antagonists of the ADORA2A pathway are being tested as adjuvants to dopaminergic drugs to treat Parkinson’s disease and schizophrenia.


Storage:

GeneBLAzer® ADORA2A CHO-K1 DA Cells and ADORA2A-CRE-bla CHO-K1 Cells are shipped on dry ice. Store in liquid nitrogen immediately upon receipt or thaw for immediate use.


Contents:
GeneBLAzer® ADORA2A 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:
•ADORA2A CHO-K1 DA cells
•LiveBLAzer™-FRET B/G Loading Kit, 70µg
•Solution D, 1ml
•2 protocols
•Certificate of Analysis
GeneBLAzer® ADORA2A 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:
•ADORA2A CHO-K1 DA cells
•1 protocol
•Certificate of Analysis
GeneBLAzer® ADORA2A-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:
•ADORA2A-CRE-bla CHO-K1 cells
•1 protocol
•Certificate of Analysis

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™ EDNRA HEK 293T DA Assay Kit

This cell line is available as dividing cells by requesting a quote for K1223 from dividingcells@invitrogen.com

GeneBLAzer® EDNRA HEK 293T DA (Division Arrested) cells and EDNRA-NFAT-bla HEK 293T cells contain the human Endothelin type A (EDNRA) receptor stably integrated into the CellSensor® NFAT-bla HEK 293T cell line. CellSensor® NFAT-bla HEK 293T cells (Cat. no.K1129) contain a beta-lactamase reporter gene under control of the Cyclic AMP Response Element (NFAT). 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 EDNRA HEK 293T DA cells and EDNRA-NFAT-bla HEK 293T cells are functionally validated for Z’-factor and EC50 concentrations of ET-1 (Figure 1). In addition, EDNRA-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.

Endothelin receptor type A (ENDRA) mediates signaling of endothelin proteins that regulate several critical biological processes, including development and function of blood vessels, production of certain hormones, and stimulation of cell growth and division. Defects in EDNRA-mediated signaling have been implicated in a wide range of diseases, including cancer, hypertension, and cardiovascular disorders.


Storage:

GeneBLAzer® EDNRA HEK 293T DA Cells and EDNRA-NFAT-bla HEK 293T Cells are shipped on dry ice. Store in liquid nitrogen immediately upon receipt or thaw for immediate use.


Contents:
GeneBLAzer® EDNRA HEK 293T 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:
•EDNRA HEK 293T DA cells
•LiveBLAzer™-FRET B/G Loading Kit, 70µg
•Solution D, 1ml
•2 protocols
•Certificate of Analysis
GeneBLAzer® EDNRA HEK 293T 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:
•EDNRA HEK 293T DA cells
•1 protocol
•Certificate of Analysis
GeneBLAzer® EDNRA-NFAT-bla HEK 293T cells
(LiveBLAzer™-FRET B/G Loading Kit, Solution D and other materials are required separately; please refer to the protocol).Includes:
•EDNRA-NFAT-bla HEK 293T cells
•1 protocol
•Certificate of Analysis

CellSensor™ DHFR/E2F-bla NIH3T3 Cell Line

The CellSensor® dhfr(E2F)-bla NIH 3T3 Cell Line contains a beta-lactamase reporter gene under control of the E2F/DP1 binding sequence from the DHFR gene promoter, stably integrated into NIH 3T3 cells. This cell line is a clonal population isolated by flow cytometry in response to 10% newborn bovine serum. This cell line has been tested for assay performance under variable conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time, and validated for Z-factor using newborn bovine serum. Additional data using alternate stimuli are shown. The G1/S cell-cycle checkpoint controls the passage of eukaryotic cells from the first gap phase (G1) into the DNA synthesis phase (S). Two cell-cycle kinases, CDK4/6-cyclin D and CDK2-cyclin E, and the transcription complex that includes Rb and E2F, are pivotal in controlling this checkpoint. During G1, the Rb-HDAC repressor complex binds to the E2F-DP1 transcription factors, inhibiting downstream transcription. Phosphorylation of Rb by CDK4/6 and CDK2 dissociates the Rb-repressor complex, permitting transcription of S-phase genes encoding for proteins that amplify the G1-to-S switch and that are required for DNA replication. Many different stimuli exert checkpoint control, including TGFβ, DNA damage, contact inhibition, replicative senescence, and growth factor withdrawal. 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™ EDNRB HEK 293T DA Assay Kit

This cell line is available as dividing cells by requesting a quote for K1224 from dividingcells@invitrogen.com

GeneBLAzer® EDNRB HEK 293T DA (Division Arrested) cells and EDNRB-NFAT-bla HEK 293T cells contain the human Endothelin type B (EDNRB) receptor stably integrated into the CellSensor® NFAT-bla HEK 293T cell line. CellSensor® NFAT-bla HEK 293T cells (Cat. no. K1129) 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 EDNRB HEK 293T DA cells and EDNRB-NFAT-bla HEK 293T cells are functionally validated for Z’-factor and EC50 concentrations of ET-1 (Figure 1). In addition, EDNRB-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.


Endothelin Receptor Type B (ENDRB) mediates signaling of endothelin proteins that regulate several critical biological processes, including the development and function of blood vessels, the production of certain hormones, and the stimulation of cell growth and division. Defects in EDNRB mediated signaling have been implicated in a wide range of diseases, including cancer, hypertension and cardiovascular disorders.



Storage:

GeneBLAzer® EDNRB HEK 293T DA Cells and EDNRB-NFAT-bla HEK 293T Cells are shipped on dry ice. Store in liquid nitrogen immediately upon receipt or thaw for immediate use.


Contents:
GeneBLAzer® EDNRB HEK 293T 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:
•EDNRB HEK 293T DA cells
•LiveBLAzer™-FRET B/G Loading Kit, 70µg
•Solution D, 1ml
•2 protocols
•Certificate of Analysis
GeneBLAzer® EDNRB HEK 293T 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:
•EDNRB HEK 293T DA cells
•1 protocol
•Certificate of Analysis
GeneBLAzer® EDNRB-NFAT-bla HEK 293T cells
(LiveBLAzer™-FRET B/G Loading Kit, Solution D and other materials are required separately; please refer to the protocol).
Includes:
•EDNRB-NFAT-bla HEK 293T cells
•1 protocol
•Certificate of Analysis

CellSensor™ p53RE-bla HCT-116 Cell Line

The CellSensor® p53RE-bla HCT-116 cell line contains a beta-lactamase reporter gene under control of the p53 response element stably integrated into HCT-116 cells. To construct this cell line, the p53RE-bla construct was transduced into HCT-116 cells by lentivirus. Flow cytometry was used to isolate cells responsive to Mitomycin. This cell line was validated for DMSO tolerance, cell number, stimulation time, substrate loading time, as well as Z'; and EC50 concentrations for Mitomycin. The CellSensor® p53RE-bla HCT-116 cell line is responsive to Mitomycin and can be used to probe p53 signaling, which is involved in DNA repair, cell cycle arrest, and in some cases, apoptosis. 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™ ADORA3-Gα15-NFAT-bla CHO-K1 cells

GeneBLAzer® ADORA3-Ga15-NFAT-bla CHO-K1 cells contain the human Adenosine A3 Receptor (ADORA3) stably integrated into the GeneBLAzer® Ga15-NFAT-bla CHO-K1 cell line. The GeneBLAzer® Ga15-NFAT-bla CHO-K1 cells (Cat. no. K1537) contain a beta-lactamase reporter gene under control of the NFAT Response Element and a promiscuous G protein, Ga15. GeneBLAzer® ADORA3-Ga15-NFAT-bla CHO-K1 cells are functionally validated for Z'-factor and EC50 concentrations of NECA.

GeneBLAzer™ LXR beta DA Assay Kit

GeneBLAzer®LXR beta DA (Division Arrested) cells and LXR beta-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human Liver-X receptor-beta (LXR 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)-LXR 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 LXR beta DA cells and LXR beta-UAS-bla HEK 293T 293 cells are functionally validated for Z' and EC50 concentrations of TO901317. In addition, LXR beta-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™ ADRB3 CHO-K1 DA Assay Kit

The GeneBLAzer® ADRB3 CHO-K1 DA (Division Arrested) cells contain the human Adrenergic Beta 3 Receptor (ADRB3) (Accession # NM_000025.1) 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 Cyclic AMP Response Element (CRE).
The ADRB3 CHO-K1 DA cells are functionally validated for Z'-factor and EC50 concentrations of Isoproterenol. In addition, ADRB3-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. 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™ 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™ OPRL1-Gqi5-NFAT-bla FreeStyle™ 293F Cells

The GeneBLAzer® OPRL1-Gqi5-NFAT-bla FreeStyle™ 293F Cell Line contains the human opioid/opiate receptor-like-1 receptor (OPRL1) stably integrated into the GeneBLAzer® Gqi5-NFAT-bla FreeStyle™ 293F Cell Line. GeneBLAzer® Gqi5-NFAT-bla FreeStyle™ 293F cells (Cat. no. K1539) contain the chimeric Gqi5 G protein and the beta-lactamase reporter gene under control of the nuclear factor of activated T cells (NFAT) response element.

OPRL1-Gqi5-NFAT-bla FreeStyle™ 293F cells have been tested for assay performance under variable conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time, and functionally validated for Z-factor and EC50 concentrations of orphanin FQ/nociceptin. These data are found in the Validation & Assay Performance Summary.

Orphanin FQ/nociceptin and OPRL1 have been implicated in various physiological functions. The most defined role is their involvement in the mediation of pain in the central nervous system and their possible roles in opiate tolerance, opiate dependence and withdrawal, and the ability to cope with stress and anxiety. Through the use of antagonists, antisense oligos, and transgenic knockout mutants, OPRL1 has been implicated in additional systems including learning, memory, and reproduction.

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

GeneBLAzer™ RXR beta DA Assay Kit

GeneBLAzer®RXR beta DA (Division Arrested) cells and RXR beta-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human Retinoid X Receptor beta (RXR 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)-RXR beta (LBD) fusion protein, the protein binds to the UAS, resulting in expression of beta-lactamase. 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 RXR beta DA cells and RXR beta-UAS-bla HEK 293T cells are functionally validated for Z' and EC50 concentrations of 9-cis retinoic acid (9-cis-RA). In addition, RXR beta-UAS-bla HEK 293T cells have been tested for assay performance under variable conditions, including DMSO concentration, cell number, and stimulation time.