Cell-Based GPCR Reporter Assays - Kits

GeneBLAzer™ EDG7-NFAT-bla HEK 293T Cells

GeneBLAzer® EDG7-NFAT-bla HEK293T cells contain the human Endothelial Differentiation Gene (EDG7) stably integrated into the CellSensor® NFAT-bla HEK293T cell line. CellSensor® NFAT-bla HEK293T cells (Cat. no. K1538) contain a beta-lactamase reporter gene under control of the Nuclear Factor of Activated T-cells (NFAT) response element.

The GeneBLAzer® EDG7-NFAT-bla HEK293T cells are functionally validated for Z' and EC50 concentrations of 18: 1 LPA. In addition, GeneBLAzer® EDG7-NFAT-bla HEK293T cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

LPA receptors were originally classified in the Endothelial Differentiation Gene (EDG) family of receptors. The EDG family was later divided into two distinct groups of receptors based on their ligand specificity for either Lysophosphatidic acid (LPA) or Sphingosine-1-Phosphate (S1P). There are three high affinity LPA receptors LPA1, LPA2, and LPA3 (formally EDG2, EDG4, and EDG7), and five high affinity S1P receptors S1P1, S1P2, S1P3, S1P4 and S1P5 (formally EDG1, EDG5, EDG3, EDG6, and EDG8). LPA receptors are differentially linked via Gi and Gq to multiple effector systems and diverse biological functions. Recent studies revealed that targeted deletion of the EDG7/LPA3 receptor results in delayed implantation and altered embryo spacing, and significantly reduced litter size in mice. These findings indicate that EDG7 plays a crucial role in implantation and mammalian reproduction.

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

GeneBLAzer™ CCKBR HEK 293T DA Assay Kit

This cell line is available as dividing cells by requesting a quote for K1231 from dividingcells@invitrogen.com

GeneBLAzer® CCKBR HEK 293T DA(Division Arrested) cells and CCKBR-NFAT-bla HEK 293T cells contain the human Cholecystokinin (CCKBR) receptor stably integrated into the CellSensor® NFAT-bla HEK 293T cell line. CellSensor® NFAT-bla HEK 293T cells (Cat. no.K1179) 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 CCKBR HEK 293T DA cells and CCKBR-NFAT-bla HEK 293T cells are functionally validated for Z’-factor and EC50 concentrations of cholecystokinin (CCK), (Figure 1). In addition, CCKBR-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.

There are two cholecystokinin receptors, referred to as A and B (CCKAR and CCKBR). Cholecystokinin (CCK) is widely distributed throughout the CNS and digestive system. Through CCKAR and CCKBR, it is thought to play a role in many digestive processes as well as anxiety, depression, psychosis, cognition and nociception. Studies have shown CCKBR could be involved in anxiety, learning and memory, as well as growth of gastric mucosa.


Storage:

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


Contents:
GeneBLAzer® CCKBR 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:
•CCKBR HEK 293T DA cells
•LiveBLAzer™-FRET B/G Loading Kit, 70µg
•Solution D, 1ml
•2 protocols
•Certificate of Analysis
GeneBLAzer® CCKBR 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:
•CCKBR HEK 293T DA cells
•1 protocol
•Certificate of Analysis

GeneBLAzer® CCKBR-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:
•CCKBR-NFAT-bla HEK 293T cells
•1 protocol
•Certificate of Analysis

Tango™ HRH3-bla U2OS Cells

The Tango™ HRH3-bla U2OS 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 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.

Tango™ CNR2-bla U2OS Cells

The 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 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.

GeneBLAzer™ BDKRB1-NFAT-bla CHO-K1 Cells

GeneBLAzer® BDKRB1-NFAT-bla CHO-K1 cells contain the human Bradykinin Receptor 1 (BDKBR1) 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® BDKRB1-NFAT-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of [Lys-des-Arg9]-Bradykinin. In addition, GeneBLAzer® BDKRB1-NFAT-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

The Bradykinin-BDKRB1 receptor has been implicated in inflammation and pain. Preclinical evidence demonstrates that interruption of BDKRB1 receptor function causes analgesia under a variety of conditions and has been well established using selective antagonists and genetically modified mice.

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

GeneBLAzer™ C5aR1-Gα15-NFAT-bla CHO-K1 Cells

GeneBLAzer® C5aR1-Gα15-NFAT-bla CHO-K1 cells contain the human Complement Component 5a Receptor 1 (C5aR1) (Accession # NM_000710) stably integrated into the GeneBLAzer® Gα15-NFAT-bla CHO-K1 cell line. GeneBLAzer® Gα15-NFAT-bla CHO-K1 (Cat. No. K1537) contains a beta-lactamase reporter gene under control of a NFAT response element and a promiscuous G Protein, Gα15, stably integrated into CHO-K1 cells.

The GeneBLAzer® C5aR1-Gα15-NFAT-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of C5a. In addition, GeneBLAzer® C5aR1-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.

GeneBLAzer™ MC2R CHO-K1 DA Assay Kit

The GeneBLAzer® MC2R CHO-K1 DA (Division Arrested) cells contain the human melanocortin-2 receptor (MC2R), (Accession # NM_000529.1) and the Melanocortin-2 receptor accessory protein (MRAP), (Accession #NM_178817.3) 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 MC2R-CHOK1 DA cells are functionally validated for Z'-factor and EC50 concentrations of ACTH 1-24. In addition, MC2R-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. 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™ GPR23-bla U2OS Cells

Tango™ GPR23-bla U2OS cells contain the human G protein-coupled receptor 23 (GPR23) 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™ GPR23-bla U2OS cells have been functionally validated for Z’ factor and a functional response to FBS (Figure 2). In addition, Tango™ GPR23-bla U2OS cells have been tested for assay performance under variable conditions.

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.

GeneBLAzer™ M1-NFAT-bla CHO-K1 Cells

GeneBLAzer® M1-NFAT-bla CHO-K1 cells contain the human Acetylcholine (muscarinic) subtype 1 receptor (M1) stably integrated into the CellSensor® NFAT-bla CHO-K1 cell line. CellSensor® NFAT-bla CHO-K1 cells (Cat. no. K1534) contain a beta-lactamase (bla) reporter gene under control of the Nuclear Factor of Activated T-cells (NFAT) response element.

M1-NFAT-bla CHO-K1 cells are functionally validated for Z'-factor and EC50 concentrations of carbachol. In addition, M1-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 brain, M1 activation mediates "slow" neuronal excitability. Cortical and hippocampal muscarinic receptors are thought to be important in the attentional aspects of cognition. The predominant receptor subtypes in these brain areas are M1, M3, and M4. Therefore, M1 is a potential target for cognition, Alzheimer's, dementia, and schizophrenia. Studies on knock-out mouse models of M1 are also beginning to reveal potential functions of the receptor. Additional information on the muscarinic receptors can be found in reviews.

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

GeneBLAzer™ Gqo5-NFAT-bla FreeStyle™ 293F Cells

GeneBLAzer® Gqo5-NFAT-bla FreeStyle™ 293F cells contain the chimeric G-protein Gqo5 stably integrated into the CellSensor® NFAT-blaFreeStyle™ 293F cell line. CellSensor® NFAT-bla FreeStyle™ 293F cells contain a beta-lactamase reporter gene under control of a stably integrated NFAT response element. Gqo5-NFAT-bla FreeStyle™ 293F is a parental cell line designed to be used for the construction of Gi/o-coupled GPCR beta-lactamase cellular assays. The Gi/o receptor of interest should be stably expressed in the cell line by expression from the selectable vector pcDNA™6.2-hygro-DEST (Cat. no. K1233). Gqo5-NFAT-bla FreeStyle™ 293F cells have been shown to respond to the agonist of a transiently expressed D2 receptor.

Tango™ CCR4-bla U2OS Cells

The Tango™ CCR4-bla U2OS cells contain the human Chemokine (C-C Motif) Receptor 4 (CCR4) 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™ CCR4-bla U2OS cells are functionally validated for Z' and EC50 concentrations of TARC. In addition, Tango™ CCR4-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™ PTGIR-CRE-bla CHO-K1 Cells

GeneBLAzer® PTGIR-CRE-bla CHO-K1 cells contain the human Prostaglandin I2 receptor (IP) (PTGIR), (Accession # NM_000960.3) 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 CRE Response Element. The GeneBLAzer® PTGIR-CRE-bla CHO-K1 cells are functionally validated for Z’-factor and EC50 concentrations of Iloprost (Figure 1). In addition, GeneBLAzer® PTGIR-CRE-bla CHO-K1 cells have been tested for assay performance under variable conditions.

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.

Tango™ GPR39-bla U2OS Cells

Tango™ GPR39-bla U2OS cells contain the orphan human G-protein Coupled Receptor 39 (GPR39) 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™ GPR39-bla U2OS cells have been functionally validated for a receptor specific response to FBS. In addition, Tango™ GPR39-bla U2OS cells have been tested for assay performance under variable conditions.