Shop All Cell Based Assay Kits

CellSensor™ CRE HEK 293T DA Assay Kit

The CellSensor® CRE HEK 293T DA (Division Arrested) cells contains a beta-lactamase (bla) reporter gene under control of a cAMP response element (CRE) stably integrated into HEK293T cells. Fluorescence activated cell sorting (FACS) was used to isolate clones responsive to stimulation of the CRE pathway by forskolin. The cell line was validated for DMSO tolerance, cell number, stimulation time, substrate loading conditions, and EC50 concentration of forskolin. 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 be used in other assays where a sensitive readout to intracellular changes to cAMP is needed.

GeneBLAzer™ RAR-gamma DA Assay Kit

GeneBLAzer®RAR gamma DA (Division Arrested) cells and RAR gamma-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human retinoic acid receptor gamma (RAR gamma) 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 gamma (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 RAR gamma DA cells and RAR gamma-UAS-bla HEK 293T cells are functionally validated for Z' and EC50 concentrations of all-trans retinoic acid (ATRA). In addition, RAR gamma-UAS-bla HEK 293T cells have been tested for assay performance under variable conditions, including DMSO concentration, cell number, and stimulation time.

Tango™ H4-bla U2OS

The Tango™ H4-bla U2OS cells contain the human Histamine Receptor 4 (H4) 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™ H4-bla U2OS cells are functionally validated for Z' and EC50 concentrations of 4-Methylhistamine. In addition, Tango™ H4-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.

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

GeneBLAzer™ T-Rex™ GPR119-CRE-bla CHO-K1 Cells

The GeneBLAzer® T-Rex™ GPR119-CRE-bla CHO-K1 cells contain the human G-Protein Receptor 119 (GPR119), (Accession # NP_848566) 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.

The T-Rex™ GPR119-CRE-bla CHO-K1 cells are functionally validated for Z'-factor and EC50 concentrations of AR231453(1). In addition, T-Rex™ GPR119-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.

Tango™ EDG4-bla U2OS Cells

The Tango™ EDG4-bla U2OS cells contain the human Endothelial Differentiation Gene 4 (EDG4) 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™ EDG4-bla U2OS DA cells have been functionally validated for Z' factor and EC50 concentrations of LPA (18:1). In addition, Tango™ EDG4-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™ 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.

CellSensor™ T-REx™ NICD CSL-bla HeLa Cell Line

The CellSensor® T-REx™ NICD CSL-bla HeLa cell line was engineered by lentiviral transduction of HeLa cervical cancer cells with a notchresponse element driving beta-lactamase reporter gene expression (CSL-bla) along with tetracycline repressor and tetracycline (or the tetracycline analog, doxycycline)-inducible NICD (notch intracellular domain) constructs. This cell line is a clonal population isolated by flow cytometry. Addition of doxycycline to these cells allows for regulated NICD transcription factor expression and subsequent beta-lactamase expression. This cell line has been tested for robust assay performance by assessing a variety of parameters, including cell plating number, DMSO tolerance, stimulation time, and substrate loading time. Assay validation was performed using serial dilutions of doxycycline. Additional testing data using RNAi and alternate stimuli are also available.

GeneBLAzer™ AR-UAS-bla Griptite™ Cells

AR-UAS-bla GripTite™ 293 cells contain the ligand-binding domain (LBD) of the rat Androgen receptor (AR) fused to the DNA-binding domain of GAL4 stably integrated in the GeneBLAzer®UAS-bla GripTite™ 293 cell line. GeneBLAzer®UAS-bla GripTite™ 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)-AR (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. AR-UAS-bla GripTite™ 293 cells are functionally validated for Z' and EC50 concentrations of R1881. 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-NFAT-bla CHO-K1 Cells

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.

LanthaScreen™ STAT5 (JAK2 V617F) U2OS Cell Line

LanthaScreen® GFP Cellular Assays allow for the analysis of post-translational modifications for a number of target proteins in an live-cell context. The JAK2⁄STAT5 signaling pathway plays an essential role in blood cell formation in response to cytokines such as GM-CSF, IL-3, and EPO. The recent discovery of an activating mutation in JAK2 (V617F) present in high percentage of myeloproliferative disease (MPD) patients suggests that this mutant JAK2 activity is a potential therapeutic target for certain forms of MPD. The assay described in this summary makes use of a cell line engineered for expression of the constitutively-active mutant kinase JAK2 V617F. By co-expressing GFP-STAT5 in this background, the phosphorylation state of STAT5 (specifically residue Tyr 694⁄699) can be modulated with JAK2 V617F inhibitors and analyzed in cell lysates using an anti-STAT5 [pTyr 694⁄699] and LanthaScreenTM terbium-anti-mouse antibody pair. GFP-STAT5 Lentivirus was transduced into U2OS cells followed by selection with Blasticidin. The selected pool was then transfected with a GST-JAK2 V617F construct using LipofectamineTM LTX, followed by selection with Geneticin. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as sorting marker, and has been screened for the constitutive expression of GFP-STAT5 and GST-JAK2 V617F. Using a lytic TR-FRET immuno-assay, this cell line is validated for IC50 and Z' under optimized conditions using JAK Inhibitor I (Pyridone 6) as a small molecule inhibitor for JAK2 V617F-mediated GFP-STAT5 phosphorylation. This assay has also been tested for assay performance under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance and lysis⁄equilibration time.

Tango™ AGTR1-bla U2OS Cells

Tango™ AGTR1-bla U2OS cells contain the human Angiotensin Receptor 1 (AGTR1) (AGTR1) 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™ AGTR1-bla U2OS cells have been functionally validated for Z’ factor and EC50 concentrations of Angiotensin II (Figure 1). In addition, Tango™ AGTR1-bla U2OS cells have been tested for assay performance under variable conditions.

CellSensor™ Myc-bla HCT116 Cell Line

The CellSensor™Myc-bla HCT116 cell line contains a beta-lactamase reporter gene under the control of Myc binding sequences. The construct was transduced into HCT116 cells using a lentiviral system. HCT116 is a colon cancer cell line which expresses a mutated form of beta-catenin. This form of beta-catenin leads to the accumulation of beta-catenin and constitutive activation of downstream genes such as Myc. This cell line is a clonal population isolated by flow cytometry. It has been validated for cell plating density and DMSO tolerance. The signaling pathway has been validated using RNAi against c-Myc and ICG-001, an inhibitor of the wnt-beta-catenin pathway. The expression of the mutant beta-catenin in HCT116 cells results in constitutive activation of beta-lactamase in this CellSensor™line, which can be knocked down by ICG-001 (Figure 1) or Myc RNAi (Figure 2).

LanthaScreen™ STAT3 GripTite™ Cell Line

The JAK/STAT3 signaling pathway is known to be activated by cytokines such as IL-6. In this pathway, binding of IL-6 to its cell-surface receptors results in the activation of JAKs, which in turn phosphoactivate STAT3 proteins at a specific tyrosine residue (Y705). LanthaScreen® STAT3 GripTite™ is a human cell line that constitutively expresses a GFP-STAT3 fusion protein. The JAK/STAT signaling pathway is known to be functionally intact in this cell line; therefore, the GFP-STAT3 fusion protein serves as a substrate for IL-6-inducible phosphorylation. Using this cell line, a lytic immunoassay has been developed in which the phosphorylation state of GFP-STAT3 is detected in cell lysates using a terbium-labeled anti-pY705-STAT3 antibody in a time-resolved FRET (TR-FRET) readout.

The GFP-STAT3 DNA expression construct was transfected into the GripTite™ HEK 293 cell line using Lipofectamine™ 2000 transfection reagent, and the transfected cells were selected with blasticidin. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as a sorting marker. The assay utilizing this cell line is validated for EC50 and Z’-factors under optimized conditions using IL-6 as a ligand for JAK-mediated GFP-STAT3 phosphorylation. This assay has also been tested under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance, and assay equilibration time. An alternate "mix-and-read" assay format is also described.

GeneBLAzer™ LXRβ-UAS-bla HEK 293T Cells

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. LXR beta-UAS-bla HEK 293T 293 cells are functionally validated for Z' and EC50 concentrations of TO901317. In addition, 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™ ERβ-UAS-bla GripTite™ Cells

ER beta-UAS-bla GripTite™ 293 cells contain the ligand-binding domain (LBD) of the human Estrogen receptor beta (ER beta) fused to the DNA-binding domain of GAL4 stably integrated in the GeneBLAzer®UAS-bla GripTite™ 293 cell line. GeneBLAzer®UAS-bla GripTite™ 293 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)-ER 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. ER beta-UAS-bla GripTite™293 cells are functionally validated for Z' and EC50 concentrations of 17-beta Estradiol. In addition, ER beta-UAS-bla GripTite™293 cells have been tested for assay performance under variable conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time. Additional testing data using alternate stimuli are also available.

GeneBLAzer™ PTGDR-CRE-bla CHO-K1 Cells

GeneBLAzer® PTGDR-CRE-bla CHO-K1 cells contain the human Prostanoid (PTGDR) Receptor 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 Cyclic AMP Response Element (CRE) response element.

The GeneBLAzer® PTGDR-CRE-bla CHO-K1cells are functionally validated for Z' and EC50 concentrations of Prostaglandin D2. In addition, GeneBLAzer® PTGDR-CRE-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

The PTGDR receptor also known as DP, is a Gs-coupled receptor which activates adenylate cyclase. PTGDR receptors are located in vascular smooth muscle cells and blood platelets, and are also believed to be expressed in the brain, where they may be involved in the regulation of sleep. In addition, knockout studies in mice suggest that PTGDR may be involved in allergic and asthmatic responses. Potent agonists and antagonists for PTGDR have been identified.

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

LanthaScreen™ PRAS40 293E Cell Line

The PI3K/AKT pathway is activated by various stimuli (including insulin and IGF-1) and mediates signals for cell growth, cell survival, translation, and inhibition of apoptosis. PRAS40 (Proline-rich AKT substrate, 40 kDa) is a cytosolic protein that is phosphorylated at position Thr246 by AKT directly upon stimulation of the pathway. PRAS40 has been shown to bind to the mTORC1 complex and inhibit downstream signaling (i.e., phosphorylation of 4E-BP1 or p70S6K). LanthaScreen® PRAS40 HEK293E is a human cell line which constitutively expresses GFP-PRAS40 fusion proteins. The kinase substrate
was introduced using lentiviral transduction followed by selection with blasticidin. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as a sorting marker. The PI3K/AKT pathway is known to be active in the highly insulin-responsive HEK293E cell background. Using this cell line, a homogenous ("addition only") immunoassay has been developed in a 384-well format. The phosphorylation of PRAS40 at Thr246 is detected in cell lysates using a terbium-labeled phosphospecific antibody in a time-resolved FRET (TR-FRET) readout. The performance of this assay has been tested using numerous experimental variables, including cell plating density, stimulation time, DMSO tolerance, and antibody incubation time. Under optimized conditions, the assay has been further validated and shows correct EC50 and IC50 values, with insulin as the primary agonist and PI-103 as the known inhibitor. Overall, the assay displays excellent statistical data (Z'-factor >0.6), high signal-to-background (response ratio), and is a robust cell-based readout of AKT signaling.

Tango™ HTR2A-bla U2OS Cells

The Tango™ HTR2A-bla U2OS cells contain the human Serotonin Type 2A receptor (HTR2A) 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™ HTR2A-bla U2OS cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ HTR2A-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™ PDCD4 HEK 293E Cell Line

The PI3K⁄AKT⁄mTOR pathway mediates signals for cell growth and survival, transcription and translation, and regulated cell death. The significance of mTOR has been underscored recently by the identification of two distinct cellular complexes: mTORC1 (includes raptor and is rapamycin sensitive) and mTORC2 (includes rictor and is insensitive to rapamycin). Activation of the pathway by insulin (or other mitogens) leads to the phosphorylation of programmed cell death protein 4 (PDCD4) via mTORC1 (indirectly through the ribosomal protein p70 S6 kinase). LanthaScreen® PDCD4 HEK293E is a human cell line which constitutively expresses GFP-PDCD4 fusion proteins. This kinase target was introduced using lipid transfection followed by selection with Blasticidin. These cells are a clonal population isolated by FACS using GFP fluorescence as a sorting marker. A homogenous immunoassay ("addition only", no wash steps) was developed in 384-well format using a TR FRET-based detection method to monitor the phosphorylation of Ser457 on GFP-PDCD4 using a terbium-labeled phosphospecific antibody. The assay has been optimized for performance under numerous experimental conditions (e.g., cell plating density, DMSO tolerance, insulin stimulation time, etc.) and displays excellent statistical data (Z' > 0.6). Moreover, this cell line has been validated with different stimuli and known inhibitors (shows correct EC50 and IC50 values) and serves as robust cellular readout for mTORC1 signaling.

Tango™ CCR1-bla U2OS DA Assay Kit

The Tango™ CCR1-bla U2OS DA cells contain the human Chemokine (C-C Motif) Receptor 1 (CCR1) 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™ CCR1-bla U2OS DA cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ CCR1-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.

Tango™ EDNRA-bla U2OS Cells

The Tango™ EDNRA-bla U2OS cells contain the human Endothelin Receptor A (EDNRA) 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™ EDNRA-bla U2OS cell line has been functionally validated for Z'-factor and EC50 concentrations of ET-1. The EDNRA 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.

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.

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.

Tango™ OPRM1-bla U2OS Cells

The Tango™ OPRM1-bla U2OS cells contain the human Opioid Receptor Mu 1 (OPRM1) 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™ OPRM1-bla U2OS cells have been functionally validated for Z' factor and EC50 concentrations of DAMGO. In addition, Tango™ OPRM1-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™ T-REx™ FOXO3 DBE-bla HeLa Cell Line

The CellSensor® T-REx™ FOXO3 DBE-bla HeLa cell line was engineered by lentiviral transduction of HeLa cervical cancer cells with a FOXO3 response element driving beta-lactamase expression (DBE-bla), along with a tetracycline repressor and tetracycline (or the tetracycline analog doxycycline)-inducible FOXO3 constructs. Addition of doxycycline to these cells allows for FOXO3 transcription factor expression and subsequent beta-lactamase expression, which in turn can be down-regulated by insulin-mediated activation of the PI3K/AKT signaling pathway which leads to phosphorylation and inactivation of FOXO3 and concomitant suppression of beta-lactamase expression. This cell line has been tested for robust assay performance by assessing a variety of assay parameters, including: cell plating number, DMSO tolerance, stimulation time, and substrate loading time. Assay validation was performed using serial dilutions of insulin to antagonize doxycycline- induced FOXO3 activity (Figure 1). Additional testing data using RNAi and alternate stimuli are also available.

CellSensor™ NFAT-bla RA-1 Cell Line

B cell receptor (BCR)-mediated signaling pathways are important for B cell proliferation and differentiation. Abnormal B cell signaling has been linked to various diseases such as lupus, lymphoma, and various immune disorders. Antigen binding to the BCR promotes the activation of several protein tyrosine kinases (PTK), which leads to phosphorylation of the BCR complex and recruitment and activation of the PTK Syk. This in turn promotes phosphorylation of PLCγ, Shc, and Vav. Additionally, the Tec family member Btk is recruited to the plasma membrane, where it is involved in activation of PLCγ. Initiation of B lymphocyte activation is dependent on the tyrosine phosphorylation-dependent formation of multimolecular effector protein complexes that activate downstream signaling pathways including PKC/Ca2+/NFAT. This cell-based assay has been thoroughly tested and validated by Invitrogen and is suitable for immediate use in screening applications. The following information illustrates the comprehensive assay testing completed and validation of assay performance under optimized conditions (Figure 1).

GeneBLAzer™ GPR54-NFAT-bla CHO-K1 Cells

GeneBLAzer® GPR54-NFAT-bla CHO-K1 cells contain the human KiSS1 receptor (GPR54) 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® GPR54-NFAT-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of Metastin. In addition, GeneBLAzer® GPR54-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 GPR54 receptor has been shown to be involved in tumor progression and metastasis, and in the development and functional regulation of the gonads.

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

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™ PPARδ-UAS-bla HEK 293T Cells

PPAR delta-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human Peroxisome Proliferator-Activated Receptor-delta (PPAR delta) 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)-PPAR delta (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. PPAR delta-UAS-bla HEK 293T cells are functionally validated for Z' and EC50 concentrations of L-165041. In addition, PPAR delta-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.

Tango™ HTR2A-bla U2OS DA Assay Kit

The Tango™ HTR2A-bla U2OS DA cells contain the human Serotonin Type 2A receptor (HTR2A) 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™ HTR2A-bla U2OS DA cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ HTR2A-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.

GeneBLAzer™ MC5R-CRE-bla CHO-K1 Cells

GeneBLAzer® MC5R-CRE-bla CHO-K1 cells contain the human Melanocortin Receptor 5 (MC5R) 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 Cyclic AMP Response Element (CRE) response element.

The GeneBLAzer® MC5R-CRE-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of NPD-α MSH In addition, GeneBLAzer® MC5R-CRE-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

MC5R is expressed in skin, muscle, lung, liver, spleen, and adrenal tissue. MC5R is Gs-coupled and binds all of the melanocortin peptides and ACTH. A recent study suggests that MC5R is involved in fat storage in the sebocyte cells of the skin. Another study in mice links MC5R to aggressive behavior. Mice deficient in MC5R acted less aggressively when treated with alpha-MSH.

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