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GeneBLAzer™ P2RY2-NFAT-bla CHO-K1 Cells

GeneBLAzer® P2RY2-NFAT-bla CHO-K1 cells contain the human Purinergic receptor P2, G protein-coupled, 2 (P2RY2) 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® P2RY2-NFAT-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of ATP. In addition, GeneBLAzer® P2RY2-NFAT-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

P2RY2 expression has been identified in smooth muscle, skeletal muscle, heart, spleen, lymphocytes, macrophages, bone marrow, lung, intestine, placenta, brain, kidney, and liver. P2RY2 has been implicated in the induction of chloride secretion in airway epithelial tissue, increases in coronary blood flow by vasodilation, and epidermal homeostasis.

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

CellSensor™ irf1-bla TF-1 Cell Line

Jak/Stat signaling pathways play essential roles in 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 TF1 Cell Line contains a beta - lactamase reporter gene under control of the irf - 1 response element stably integrated into TF1 cells. TF1 cells are a human erythroleukemia cell line that is growth - dependent on GM - CSF and have an intact GM - CSF - JAK2 - STAT5 pathway. This cell line is validated for EC50 and Z’ - factor using GM - CSF. This cell line has also been tested under variable experimental conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time. Responsiveness to EPO and Jak Inhibitor 1 was also tested (Figures 2 and 3). CellSensor® irf1 - bla TF1 cells were stimulated in triplicate with GM - CSF over the indicated concentration range in a 384 - well format. Cells were incubated for 5 hrs with agonist and 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 were plotted for each replicate against the indicated concentrations of GM - CSF. CellSensor® irf1 - bla TF1 cells were stimulated with Epo over the indicated concentration range in a 384 - well format. Cells were incubated for 5 hrs with agonist and 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 Response Ratios were plotted against the indicated concentrations of Epo. CellSensor® irf1 - bla TF1 cells were treated with Jak Inhibitor 1 over the indicated concentration range in a 384 - well format. Cells were then stimulated with GM - CSF or EPO for 5 hrs 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. These values were converted to percent inhibition relative to a set of controls (unstimulated cells and EC80 of GM - CSF or EPO treated cells) and plotted against the indicated concentrations of Jak Inhibitor 1. Academic and non-profit customers, please inquire for special pricing.

Tango™ CCR1-bla U2OS Cells

The Tango™ CCR1 -bla U2OS 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 cells have been functionally validated for Z' factor and EC50 concentrations of established ligands. In addition, Tango™ CCR1-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.

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™ TR beta DA Assay Kit

GeneBLAzer®TR beta DA(Division Arrested) cells and 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. 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 beta DA cells and 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™ M1 CHO-K1 DA Assay Kit

This cell line is available as dividing cells by requesting a quote for K1230 from dividingcells@invitrogen.com

GeneBLAzer® M1 CHO-K1 DA(Division Arrested) cells and 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.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 M1 CHO-K1 DA cells and M1-NFAT-bla CHO-K1 cells are functionally validated for Z’-factor and EC50 concentrations of carbachol, (Figure 1). In addition, M1-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.

The Musacarinic 1(M1) Receptor is localized principally in the cortical and hippocampal regions of the brain and has been implicated in various neurological conditions including depression, anxiety, and sleep disorders.


Storage:

GeneBLAzer® M1 CHO-K1 DA Cells and M3-NFAT-bla CHO-K1 Cells are shipped on dry ice. Store in liquid nitrogen immediately upon receipt or thaw for immediate use.


Contents:
GeneBLAzer® M1 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:
•M1 CHO-K1 DA cells
•LiveBLAzer™-FRET B/G Loading Kit, 70µg
•Solution D, 1ml
•2 protocols
•Certificate of Analysis
GeneBLAzer® M1 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:
•M1 CHO-K1 DA cells
•1 protocol
•Certificate of Analysis

GeneBLAzer® M1-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:
•M1-NFAT-bla CHO-K1 cells
•1 protocol
•Certificate of Analysis

CellSensor™ SIE-bla HEK293T Cell Line

The CellSensor® SIE-bla HEK 293T cell line contains a beta-lactamase reporter gene under control of the SIE response element stably integrated into HEK 293T cells. The cell line was created through sorting by FACS of cells responsive to stimulation of the SIE pathway with interleukin 6 (IL-6). The cell line was validated for DMSO tolerance, incubation time with stimulant, and substrate loading conditions. The CellSensor® SIE-bla HEK 293T cell line responds to agonist treatment as expected from literature and can be adapted for high throughput screening for agonists or antagonists of the SIE pathway with compound libraries. Candidate drugs can also be tested for dose response against this cell line. Academic and non-profit customers, please inquire for special pricing.

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.

Tango™ MCHR2-bla U2OS Cells

The Tango™ MCHR2-bla U2OS cells contain the human Melanin-concentrating hormone receptor 2 (MCHR2) 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™ MCHR2-bla U2OS cells are functionally validated for Z' and EC50 concentrations of I-TAC (CXCL11). In addition, Tango™ MCHR2-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™ ADRB1-CRE-bla CHO-K1 Cells

The GeneBLAzer ® ADRB1-CRE-bla CHO-K1 cells contain the human Adrenergic Beta-1 Receptor (ADRB1), (Accession # NM_000684) 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 ADRB1-CRE-bla CHO-K1 cells are functionally validated for Z'-factor and EC50 concentrations of (-) Denopamine. In addition, ADRB1-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™ M4-Gqo5-NFAT-bla CHO-K1 Cells

GeneBLAzer® M4 Gqo5-NFAT-bla CHO-K1 cells contain the human Acetylcholine (muscarinic) subtype 4 receptor (M4) stably integrated into the GeneBLAzer® Gqo5-NFAT-bla CHO-K1 cell line. GeneBLAzer® Gqo5-NFAT-bla CHO-K1 cells (Cat. no. K1536) contain a beta-lactamase reporter gene under control of the Nuclear Factor of Activated T-cells (NFAT) response element and the Chimeric G protein, Gqo5.

The GeneBLAzer® M4 Gqo5-NFAT-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of Cabachol. In addition, GeneBLAzer® M4 Gqo5-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 clinical implications of the M4 receptor are unknown, however studies with knockout mice suggest that M4 may have implications in dopaminergic function, karatinocyte migration and wound healing, anxiolysis, and analgesia. In addition, M4 may be important in memory circuits and have implications in Alzheimer's Disease. Additional information on the muscarinic receptors can be found in literature.

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

GeneBLAzer™ ADORA2A-CRE-bla CHO-K1 Cells

GeneBLAzer® ADORA2A-CRE-bla CHO-K1 cells contain the human Adenosine Receptor A2A (ADORA2A) 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® ADORA2A-CRE-bla CHO-K1 cells are functionally validated for Z' and EC50 concentrations of NECA. In addition, GeneBLAzer® ADORA2A-CRE-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary.

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.

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

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.

Tango™ CHRM2-bla U2OS Cells

The Tango™ CHRM2-bla U2OS cells contain the human Cholinergic muscarinic 2 receptor (CHRM2) 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™ CHRM2-bla U2OS cells have been functionally validated for Z' factor and EC50 concentrations of Carbachol. In addition, Tango™ CHRM2-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™ PAC1-CRE-bla CHO-K1 Cells

The GeneBLAzer® PAC1-CRE-bla CHO-K1 cells contain the human Adenylate Cyclase Activating Polypeptide 1 Receptor 1 (ADCYAP1R⁄PAC1) 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 PAC1-CRE-bla CHO-K1 cells are functionally validated for Z'-factor and EC50 concentrations of established ligands. In addition, PAC1-CRE-bla CHO-K1 cells have been tested for assay performance under variable conditions. These data are found in the Validation & Assay Performance Summary