Screen Quest™ HEK-CNGC-Chemokine (C-X-C motif) Receptor 4 Cells
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Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
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Shipping | Standard overnight for United States, inquire for international |
Storage, safety and handling
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
Storage | Liquid nitrogen |
UNSPSC | 12352200 |
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Overview | ![]() ![]() |
G protein coupled receptors (GPCRs) are one of the largest receptor classes targeted by drug discovery programs. Calcium flux (coupled via Gq pathway) assay is a preferred method in drug discovery for screening GPCR targets. However, over 60% of the known GPCRs signal through adenylyl cyclase activity coupled to cAMP. Most of the existing cAMP assays not only require cell lysis but also lack both temporal and spatial resolution. Screen Quest™ cell lines enable to investigate GPCRs that do not conventionally couple through intracellular calcium. Screen Quest™ cell lines are based on a series of G-protein chimeras, including the promiscuous G-protein, Gα16, and an exogenous cyclic nucleotide-gated channel (CNGC). The chimeras consist of the alpha subunit of a Gq-protein complex whose 5 carboxy-terminal amino acids have been replaced with those from one of the other G-proteins (either Gαs, Gαi, Gαo, or Gαz). These amino acids are responsible for the coupling of the receptor to its G-protein. Co-expression of these chimeras or CNGC with specific non-Gq-coupled receptors may result in the generation of an intracellular calcium signal upon receptor stimulation. Screen Quest™ HEK-CNGC-Chemokine (C-X-C motif) Receptor 4 (CXCR4) cell line is HEK-293 cells stably transfected with both the CNGC and human Chemokine (C-X-C motif) Receptor 4. The constitutively expressed CNGC in the cells responds in real-time to increases or decreases in intracellular cAMP levels by coordinately altering cation flux (e.g., calcium, potassium or sodium). Activation of the CXCR4 in these cells by specific ligands can be detected with either a calcium-sensitive fluorescent indicator (such as Calbryte 520 AM, Cal-520 AM, Fluo-8 AM, or Fluo-4 AM and the corresponding no wash calcium kit) or an AAT’s optimized membrane-potential assay kit. This cell line has been successfully used in drug discovery and screening environments for studying GPCRs that do not conventionally couple through intracellular calcium. It has been effectively used with FLIPR and FDSS systems.
Images

Figure 2. Figure 1. Response of HEK-CXCR4 cells and parental cells to SDF-1α.
Both CXCR4 cells and parental cells were plated overnight in 20 µL/well culture medium in a Poly-D-Lysine coated black wall/clear bottom 384-well plate. The cells were incubated with equal volume (20 µL) of MP dye working solution at room temperature for 2 hours. Two readings were obtained prior to and 25 min after the addition of SDF-1α. Ratios of the two readings (F/F0) are plotted in the figure.
A. Dose response curve of SDF-1α in HEK-CXCR4 cell line. EC50 = 14 ng/ml in the presence of PDE inhibitor Ro20-1724 and β-adrenoceptor agonist isoproterenol
B: Parental cells do not respond to SDF-1α.
Both CXCR4 cells and parental cells were plated overnight in 20 µL/well culture medium in a Poly-D-Lysine coated black wall/clear bottom 384-well plate. The cells were incubated with equal volume (20 µL) of MP dye working solution at room temperature for 2 hours. Two readings were obtained prior to and 25 min after the addition of SDF-1α. Ratios of the two readings (F/F0) are plotted in the figure.
A. Dose response curve of SDF-1α in HEK-CXCR4 cell line. EC50 = 14 ng/ml in the presence of PDE inhibitor Ro20-1724 and β-adrenoceptor agonist isoproterenol
B: Parental cells do not respond to SDF-1α.
Citations
View all 2 citations: Citation Explorer
Activation of P2X7 and P2Y11 purinergic receptors inhibits migration and normalizes tumor-derived endothelial cells via cAMP signaling
Authors: Avanzato, D and Genova, T and Pla, A Fiorio and Bernardini, M and Bianco, S and Bussolati, B and Mancardi, D and Giraudo, E and Maione, F and Cassoni, P and others, undefined
Journal: Scientific Reports (2016)
Authors: Avanzato, D and Genova, T and Pla, A Fiorio and Bernardini, M and Bianco, S and Bussolati, B and Mancardi, D and Giraudo, E and Maione, F and Cassoni, P and others, undefined
Journal: Scientific Reports (2016)
The M2 muscarinic receptors are essential for signaling in the heart left ventricle during restraint stress in mice
Authors: Tomankova, Hana and Valuskova, Paulina and Varejkova, Eva and Rotkova, Jana and Benes, Jan and Myslivecek, Jaromir
Journal: Stress (2015)
Authors: Tomankova, Hana and Valuskova, Paulina and Varejkova, Eva and Rotkova, Jana and Benes, Jan and Myslivecek, Jaromir
Journal: Stress (2015)
References
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Cardiac Hypertrophy Is Inhibited by a Local Pool of cAMP Regulated by Phosphodiesterase 2
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cAMP controls the balance of the propulsive forces generated by the two flagella of Chlamydomonas
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Application notes
A Comparison of Fluorescent Red Calcium Indicators for Detecting Intracellular Calcium Mobilization in CHO Cells
A Meta-Analysis of Common Calcium Indicators
A New Red Fluorescent & Robust Screen Quest™ Rhod-4™ Ca2+Indicator for Screening GPCR & Ca2+ Channel Targets
A New Robust No-Wash FLIPR Calcium Assay Kit for Screening GPCR and Calcium Channel Targets
A Novel NO Wash Probeniceid-Free Calcium Assay for Functional Analysis of GPCR and Calcium Channel Targets
A Meta-Analysis of Common Calcium Indicators
A New Red Fluorescent & Robust Screen Quest™ Rhod-4™ Ca2+Indicator for Screening GPCR & Ca2+ Channel Targets
A New Robust No-Wash FLIPR Calcium Assay Kit for Screening GPCR and Calcium Channel Targets
A Novel NO Wash Probeniceid-Free Calcium Assay for Functional Analysis of GPCR and Calcium Channel Targets
FAQ
Are there any calcium indicators that don't require probenecid (PBC)?
Are there upgraded trypan blue derivatives for cell viability testing?
Can I intracellularly measure mitochondria calcium flux and changes in mitochondria membrane potential at the same time?
Do you offer any products for measuring intracellular calcium concentration or movement by flow cytometry?
Does EDTA inactivate proteinase K?
Are there upgraded trypan blue derivatives for cell viability testing?
Can I intracellularly measure mitochondria calcium flux and changes in mitochondria membrane potential at the same time?
Do you offer any products for measuring intracellular calcium concentration or movement by flow cytometry?
Does EDTA inactivate proteinase K?