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GeneBLAzer™ TRα-UAS-bla HEK 293T Assay Kit

TR alpha-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human Thyroid hormone receptor alpha(TR 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)-TR alpha (LBD) fusion protein, the protein binds to the UAS, resulting in expression of beta-lactamase. TR alpha-UASbla HEK 293T 293 cells are functionally validated for Z' and EC50 concentrations of T3 Thryoid hormone. In addition, TR 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™ PPAR alpha UAS-bla HEK 293T Cellular Assay Kit

The GeneBLAzer® PPAR alpha-UAS-bla HEK 293T Cellular Assay provides an accurate, sensitive, and easy-to-use method for monitoring the cellular response of PPAR alpha to drug candidates or other stimuli.

Features of the The GeneBLAzer® PPAR alpha-UAS-bla HEK 293T Cellular Assay:
Convenience—plate cells and perform assay without additional cell culture steps
Low Noise—the kit uses a Fluorescence Resonance Energy Transfer (FRET)-based ratiometric readout for minimum noise and error
Low Background—the GAL4 fusion prevents any background from endogenous receptor

The Cells
The GeneBLAzer® PPAR alpha-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human peroxisome proliferator-activated receptor alpha (PPAR alpha) fused to the DNA-binding domain of GAL4 transiently transduced (via BacMam virus) into the GeneBLAzer® UAS-bla HEK293T cell line. GeneBLAzer® UAS-bla HEK 293T cells stably express a β-lactamase reporter gene under the transcriptional control of an upstream activator sequence (UAS).

The Assay
When an agonist binds to the LBD of the GAL4 (DBD)-PPAR alpha (LBD) fusion protein, the protein binds to the UAS, resulting in expression of β-lactamase. The FRET ratiometric readout from the live-cell substrate reduces the absolute and relative errors that can mask the underlying biological response of interest. PPAR alpha UAS-bla HEK 293T cells have been tested for assay performance using variable assay conditions, including cell number, stimulation time, substrate loading time and have been validated for Z′ and EC50 concentrations of GW7647. Additional testing data using alternate stimuli are also available. See the "validation packet" in the documents below to view the data.

Speed and Convenience
Because PPAR alpha UAS-bla HEK 293T cells are transiently transduced with BacMam virus, they are ready to be used in an assay without any additional cell culture steps. Just plate the cells and assay as outlined in the protocol.

Please email our Custom Services team to inquire about obtaining these cells at economical bulk scales.

GeneBLAzer™ TR alpha DA Assay Kit

GeneBLAzer®TR alpha DA(Division Arrested) cells and TR alpha-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human Thyroid hormone receptor alpha(TR 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)-TR 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 TR alpha DA cells and TR alpha-UASbla HEK 293T 293 cells are functionally validated for Z' and EC50 concentrations of T3 Thryoid hormone. In addition, TR 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™ RARα-UAS-bla HEK 293T Cells

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 betalactamase. 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.

GeneBLAzer™ PR DA Assay Kit

GeneBLAzer®PR DA (Division Arrested) cells and PR-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human Progesterone Receptor (PR) 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)-PR (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 PR DA cells and PR UAS- bla HEK 293T cells are functionally validated for Z' and EC50 concentrations of R5020. In addition, PR-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.

GeneBLAzer™ PPAR gamma DA Assay Kit

GeneBLAzer®PPAR gamma DA (Division Arrested) cells and PPAR gamma-UAS-bla HEK 293H cells contain the ligand-binding domain (LBD) of the human Peroxisome Proliferator-Activated Receptor-gamma (PPAR gamma) fused to the DNA-binding domain of GAL4 stably integrated in the GeneBLAzer®UAS-bla HEK 293H cell line. GeneBLAzer®UAS-bla HEK 293H 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 gamma (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 PPAR gamma DA cells and PPAR gamma-UAS-bla HEK 293H cells are functionally validated for Z' and EC50 concentrations of Rosiglitazone. In addition, PPAR gamma-UAS-bla HEK 293H cells have been tested for assay performance under variable conditions, including DMSO concentration, cell number, and stimulation time.

GeneBLAzer™ MR DA Assay Kit

GeneBLAzer®MR DA(Division Arrested) cells and MR-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human Mineralcorticoid Receptor (MR) fused to the NAbinding 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. 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 MR DA cells and MRUAS-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.

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™ RARγ-UAS-bla HEK 293T Cells

The GeneBLAzer® 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 (Cat. no. K1104) 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, it binds to the UAS, inducing transcription of beta-lactamase. GeneBLAzer® RAR gamma-UAS-bla HEK 293T cells have been tested for assay performance using variable assay conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time. The Z´ and EC50 concentrations of all-trans-retinoic acid and other stimuli have been determined. Additional testing data are available upon request. RAR gamma is a member of the retinoic acid receptor family of nuclear receptors. Retinoids, vitamin A and its natural and synthetic analogues, are a very important group of hormones that regulate a wide variety of biological functions including embryogenesis, cell growth, and cell differentiation. Retinoids are beneficial in a wide variety of clinical cases including dermatology, oncology, diabetes, and diseases associated with HPV. RAR gamma is the predominant RAR subtype present in the skin and it constitutes more than 90% of the total skin or keratinocyte RAR repertoire. Skin irritation is a classical toxicity associated with RAR pan-agonists, like RA or TTNPB, and is believed to be a manifestation of RAR gamma activation. Furthermore, it has been shown that tumor-specific apoptosis can be driven by RAR gamma selective agonists, for pancreatic cancer cells.

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.

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.

GeneBLAzer™ ERRα-UAS-bla HEK 293T Cells

ERR alpha-UAS-bla HEK 293T cells contain the ligand-binding domain (LBD) of the human Estrogen related receptor alpha (ERR 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)-ERR 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. ERR alpha-UAS-bla HEK 293T cells are functionally validated for Z' and EC50 concentrations of XCT790, an inverse agonist. In addition, ERR alpha-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.

GeneBLAzer™ ER Beta DA Assay Kit

GeneBLAzer®ER beta DA(Division Arrested) cells and 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. 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 ER beta DA cells and 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™ 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.

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.