CellSensor Cellular Pathway Assays

CellSensor™ AP-1-bla ME-180 Cell Line

The CellSensor® AP-1-bla ME-180 Cell Line contains a beta-lactamase reporter gene under control of the AP-1 response element stably integrated into ME-180 cells. The cell line was created through FACS sorting of cells responsive to stimulation of the AP-1 pathway with Epidermal Growth Factor (EGF). This cell line was validated for DMSO tolerance, stimulant incubation time, substrate loading conditions, Z' and EC50 concentration of Epidermal Growth Factor. The AP-1-bla ME-180 Cell Line responds to agonist treatment as expected from the literature and can be adapted for high-throughput screening for agonists or antagonists of the AP-1 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.

CellSensor™ NFκB-bla THP-1 Cell Line

The CellSensor® NFκB-bla THP-1 cell line contains a beta-lactamase reporter gene under the control of the Nuclear Factor kappa B (NFκB) response element stably integrated into THP-1 cells. To obtain the cell line, the NFκB-bla construct was transduced into THP-1 cells by lentivirus. Subsequent flow cytometry was used to isolate cells responsive to Tumor Necrosis Factor alpha (TNFα). The cell line was validated for DMSO tolerance, cell number, stimulation time, substrate loading time, and EC50 concentration of TNF&alpha. The CellSensor® NFκB-bla THP-1 cell line is responsive to TNFα and can be used to probe NFκB signaling pathways, including those involved in apoptosis, viral defense, cancer, inflammation, and autoimmune disease. 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.

CellSensor™ NFKB-bla Freestyle™ HEK293F Cell Line

Nuclear Factor kappaB (NFκB) signaling regulates genes involved in apoptosis, viral defense, cancer, inflammation, and autoimmune disease. The CellSensor® NFκB-bla FreeStyle™ 293F Cell Line is responsive to tumor necrosis factor alpha (TNFα) and can be used to probe the NFκB signaling pathway. This cell line was validated for DMSO tolerance, incubation time with stimulant and substrate loading conditions. The CellSensor® NFκB-bla FreeStyle™ 293F Cell Line contains a beta-lactamase reporter gene under the control of NFκB response element. The construct was transduced into FreeStyle™ 293F cells by lentivirus. This cell line is a pool isolated for response to TNFα by flow cytometry. It responds to agonist treatment as expected from literature. Academic and non-profit customers, please inquire for special pricing.

CellSensor™ NFKB-bla HEK293T Cell Line

The CellSensor® NFκB-bla HEK 293T cell line contains a beta-lactamase reporter gene under the control of the Nuclear Factor kappa B (NFκB) response element. To obtain the cell line, the NFκB-bla construct was transduced into HEK 293T cells by lentivirus. Subsequent flow cytometry was used to isolate clones responsive to Tumor Necrosis Factor alpha (TNFα). The cell line has been validated for DMSO tolerance, stimulation time, substrate loading time, and EC50 concentration of TNF&alpha. This cell line is responsive to TNFα and can be used to probe NFκB signaling pathways, including those that are involved in apoptosis, viral defense, cancer, inflammation, and autoimmune disease. Academic and non-profit customers, please inquire for special pricing.

CellSensor™ NFκB-bla RA 1 Cell Line

The CellSensor® NFκB-bla RA-1 cell line contains a beta-lactamase reporter gene under control of the NFκB response element stably integrated into Ramos 1 (RA-1) cells. To construct this cell line, the NFκB -bla construct was transduced into RA-1 cells by lentivirus. Flow cytometry was used to isolate cells responsive to Tumor Necrosis Factor (TNFThis cell line was validated for DMSO tolerance, cell number, stimulation time, substrate loading time, as well as Z´ and EC50 concentrations for TNF. The CellSensor® NFκB-bla RA-1 cell line is responsive to TNFand can be used to probe NFκB signaling pathways, which are involved in the regulation of apoptosis, viral replication, tumorigenesis, inflammation, and various autoimmune diseases. 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.

CellSensor™ irf1-bla CTLL-2 Cell Line

Jak/Stat signaling pathways play essential roles in the 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 CTLL2 Cell Line contains a beta - lactamase reporter gene under control of the irf - 1 response element stably integrated into CTLL2 cells. CTLL2 cells are a clone of cytotoxic T cells derived from a C57BL/6 mouse, and are cell - growth dependent on mouse Interleukin - 2 (mIL - 2). This cell line is validated for EC50 and Z' - factor under optimized conditions using mIL - 2. This cell line has also been tested under variable assay conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time. Responsiveness to pathway inhibitors was also tested. CellSensor® irf1 - bla CTLL2 cells were stimulated with mIL - 2 in triplicate over the indicated concentration range in a 384 - well format. Cells were incubated for 5 hrs with agonist 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. The 460/530 ratios for each replicate plotted against the indicated concentrations of mIL - 2. CellSensor® irf1 - bla CTLL2 cells were treated with Jak Inhibitor 1 over the indicated concentration range in a 384 - well format for 30 min. Cells were then incubated for 5 hrs with mIL - 2 agonist (1 ng/ml) in 0.5% DMSO and then combined with LiveBLAzer™ - FRET B/G Substrate (CCF4 - AM) for 2 hrs. Fluorescence emission values at 460 nm and 530 nm were obtained using a standard fluorescence plate reader, converted to percent inhibition relative to a set of controls (unstimulated cells and EC80 stimulated cells), and plotted against the indicated concentrations of Jak Inhibitor 1. Academic and non-profit customers, please inquire for special pricing.

CellSensor™ NFKB-bla Jurkat Cell Line

The CellSensor® NFκB-bla Jurkat cell line contains a beta-lactamase reporter gene under control of the Nuclear Factor Kappa B (NFκB) response element stably integrated into Jurkat cells. To construct this cell line, the NFκB-bla construct was transduced into Jurkat cells by lentivirus. Flow cytometry was used to isolate cells responsive to Tumor Necrosis Factor alpha (TNFα). This cell line was validated for DMSO tolerance, cell number, stimulation time, substrate loading time, and Z;-factor and EC50 concentrations for TNF&alpha. The CellSensor® NFκB-bla Jurkat cell line is responsive to TNFα and can be used to probe NFκB signaling pathways, including those involved in apoptosis, viral defense, cancer, inflammation, and autoimmune disease. 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.

CellSensor™ irf1-bla Ba/F3 Cell Line

Jak/Stat signaling pathways play essential roles in the 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 BA/F3 Cell Line contains a beta - lactamase reporter gene under control of the irf - 1 response element stably integrated into BA/F3 cells. This cell line is validated for EC50 and Z' - factor under optimized conditions using mouse IL - 3 (mIL - 3). This cell line has also been tested under variable experimental conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time. Responsiveness to Jak Inhibitor 1, a small molecule inhibitor, was also tested. CellSensor® irf1 - bla BA/F3 cells were treated with mIL - 3 in triplicate over the indicated concentration range in a 384 - well format. Cells were incubated for 5 hrs with mIL - 3 agonist 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. The Response Ratios for each replicate were plotted against the indicated concentrations of mIL - 3. CellSensor ® irf1 - bla BA/F3 cells were treated with Jak Inhibitor 1 for 30 min over the indicated concentration range in a 384 - well format. Cells were then incubated with mIL - 3 agonist in 0.5% DMSO for 5 hrs 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 emission ratios were plotted against the indicated concentrations of Jak Inhibitor 1. Academic and non-profit customers, please inquire for special pricing.

CellSensor™ HRE-bla ME-180 Cell Line

The CellSensor® HRE-bla ME-180 cell line contains a beta-lactamase reporter gene under control of the hypoxia response element (HRE) stably integrated into ME-180 cells. To construct this cell line, the HRE-bla construct was transduced into ME-180 cells by lentivirus. Flow cytometry was used to isolate cells responsive to deferoxamine. This cell line is validated for DMSO tolerance, cell number, stimulation time, substrate loading time, as well as Z' and EC50 concentrations for cobalt chloride and deferoxamine. The CellSensor® HRE-bla ME-180 cell line is responsive to deferoxamine and cobalt chloride and can be used to probe the hypoxia signaling pathway. This cell line can be adapted for high-throughput screening of agonist or antagonist 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.

CellSensor™ irf1-bla HEL Cell Line

Jak/Stat signaling pathways play essential roles in the cellular responses to distinct cytokines. One of 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 knock-out causes embryonic lethality due to defective erythropoiesis, suggesting the Jak2/Stat5 pathway plays important role in red blood cell formation. Recent discovery of activating mutation in JAK2 (V617F) present in high percentage of myeloproliferative disease (MPD) patients suggests Jak2/Stat5 pathway to be the potential therapeutic target for certain forms of MPD. The activated transcription factor Stat5 dimers recognize and bind 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 HEL cell line contains a beta-lactamase reporter gene under control of the interferon regulatory factor-1 (irf1) response element stably integrated into HEL cells. HEL cells are a human erythroleukemia cell line that is growth factor independent and contains a endogenous homozygous JAK2V617F mutation. This cell line validated for IC50 and Z'-Factor under optimized conditions using Jak Inhibitor 1 (Figure 1). This cell line has also been tested under variable experimental conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time. Academic and non-profit customers, please inquire for special pricing.

CellSensor™ CRE-bla Jurkat Cell Line

The CRE-bla Jurkat cell line contains a beta-lactamase reporter gene under control of a cAMP response element (CRE) stably integrated into Jurkat cells. This cell line can be used as a parental cell line to build specific G-protein coupled receptor (GPCR) assays after transfection of additional genes of interest or to detect changes in intracellular cAMP levels. CRE-bla Jurkat cells have been shown to be responsive to forskolin stimulation. Academic and non-profit customers, please inquire for special pricing.

CellSensor™ c-Fos-bla ME-180 Cell Line

The CellSensor® c-fos-bla ME-180 cell line contains a beta-lactamase reporter gene under control of the c-fos response element stably integrated into ME-180 cells. To construct this cell line, the c-fos-bla construct was transduced into ME-180 cells by lentivirus. Flow cytometry was used to isolate cells responsive to epidermal growth factor (EGF). This cell line is validated for DMSO tolerance, cell number, stimulation time, substrate loading time, as well as Z' and EC50 concentrations for EGF. The CellSensor® c-fos-bla ME-180 cell line is responsive to EGF, HGF, IL-6, OSM, TNFα, IL-1-alpha; and PMA/Thaps, and can be used to probe the JNK/P38/MAPK and JAK/STAT signaling pathways. This cell line can be adapted for high-throughput screening of agonist or antagonist 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.

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.

CellSensor™ NFkB-bla ME-180 Cell Line

The CellSensor® NFκB-bla ME-180 cell line contains a beta-lactamase reporter gene under control of the Nuclear Factor Kappa B (NFκB) response element stably integrated into ME-180 cells. The cell line was created through fluorescence activated cell sorting (FACS) of cells responsive to stimulation of the NFκB pathway with tumor necrosis factor alpha (TNFα). The cell line was validated for DMSO tolerance, incubation time with stimulant, and substrate loading conditions. The CellSensor® NFκB-bla ME-180 cell line responds to agonist treatment as expected from literature and can be adapted for high-throughput screening for agonists or antagonists of the NFκB 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.

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.