Name: Screen Quest™ Live Cell cAMP Assay Service Pack
Brand: AAT Bioquest
SKU: 36382
Availability: InStock

Screen Quest™ Live Cell cAMP Assay Service Pack

Screen Quest™ Live Cell cAMP Assay Service Pack is a live-cell fluorescence assay reagent pack for monitoring activation of GPCRs through Gα16-mediated calcium mobilization.

Gα16-mediated receptor coupling: Enables selected non-Gq-coupled GPCRs to generate a calcium response upon receptor activation
Live-cell fluorescence readout: Uses Calbryte™ 520 NW to monitor intracellular calcium changes without cell lysis
Integrated assay reagents: Includes Gα16 DNA, Transfectamine™ 5000, and calcium detection reagents
Research applications: Suitable for functional screening of compatible Gs- and Gi-coupled GPCRs using a redirected calcium readout

Vasopressin responses in CHO cells. CHO cells were transiently transfected with Ga16 and vasopressin receptor 2 (V2R). CHO cells were cultured in a 6-well plate and grown to ~60% confluence. Equal amounts of Ga16 (1.5 µg) and V2R plasmids (1.5 µg) were transfected with 9 µL of Transfectamine™ 5000. Cells were transferred to a 96-well plate at 50,000 cells/100 µL/well ~ 30 hours after transfection. 100 µL of Calbryte™ 520NW dye-loading solution was added ~ 48 hours after transfection and incubated at 37 °C for 45 minutes. Vasopressin (50 µL/well) was added using FlexStation 3 to achieve the final concentration of 100 nM.

Protocol

Catalog	Size	Price
36382	100 Tests	Price
36383	200 Tests	Price
36384	1000 Tests	Price

Overview

G protein coupled receptors (GPCR) 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™ Live Cell cAMP Assay Service Pack provides the real-time monitoring of intracellular cAMP change in a high-throughput format without a cell lysis step. The assay works through the cell lines that contain either an exogenous cyclic nucleotide-gated channel (CNGC) or the promiscuous G-protein, Gα16. The channel is activated by elevated levels of intracellular cAMP, resulting in ion flux and cell membrane depolarization which can be detected with either a fluorescent calcium (such as Calbryte 520 AM, Cal-520® AM, Fluo-8® AM, or Fluo-4 AM and corresponding no wash calcium kits) or a fluorescent membrane potential dye. Co-expression of Gα16 with specific non-Gq-coupled receptors will result in the generation of an intracellular calcium signal upon receptor stimulation. The Screen Quest™ Live Cell cAMP Assay Service Pack provides all the reagents needed for the measurement of intracellular cAMP changes with a FLIPR, a FDSS or other equivalent fluorescence microplate readers. It has been successfully used to measure Gs and Gi coupled GPCR activity.

Components

Catalog 36382 (100 Tests)

Component A: Ga16 DNA	1 vial (15 µg)
Component B: Transfectamine™ 5000	1 vial (75 µL)
Component C: Calbryte™ 520 NW	1 vial
Component D: 10X Pluronic® F127 Plus	1 bottle (1 mL)
Component E: HHBS	1 bottle (9 mL)
Component F: DMSO	1 vial (100 µL)

Catalog 36383 (200 Tests)

Component A: Ga16 DNA	2 vials (15 µg/vial)
Component B: Transfectamine™ 5000	1 vial (150 µL)
Component C: Calbryte™ 520 NW	2 vials
Component D: 10X Pluronic® F127 Plus	2 bottles (1 mL/bottle)
Component E: HHBS	2 bottles (9 mL/bottle)
Component F: DMSO	1 vial (100 µL)

Catalog 36384 (1000 Tests)

Component A: Ga16 DNA	1 vial (150 µg)
Component B: Transfectamine™ 5000	1 vial (750 µL)
Component C: Calbryte™ 520 NW	1 vial
Component D: 10X Pluronic® F127 Plus	2 bottle (10 mL)
Component E: HHBS	1 bottle (100 mL)
Component F: DMSO	1 vial (250 µL)

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)

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)

References

View all 132 references: Citation Explorer

cAMP-Induced Histones H3 Dephosphorylation Is Independent of PKA and MAP Kinase Activations and Correlates With mTOR Inactivation

Authors:

Rodriguez P, Rojas J.

Journal:

J Cell Biochem (2016): 741

Changes in the Arabidopsis thaliana Proteome Implicate cAMP in Biotic and Abiotic Stress Responses and Changes in Energy Metabolism

Authors:

Alqurashi M, Gehring C, Marondedze C.

Journal:

Int J Mol Sci (2016): 852

Role of the cAMP Pathway in Glucose and Lipid Metabolism

Authors:

Ravnskjaer K, Madiraju A, Montminy M.

Journal:

Handb Exp Pharmacol (2016): 29

Odor-induced cAMP production in Drosophila melanogaster olfactory sensory neurons

Authors:

Miazzi F, Hansson BS, Wicher D.

Journal:

J Exp Biol (2016): 1798

A cardiac mitochondrial cAMP signaling pathway regulates calcium accumulation, permeability transition and cell death

Authors:

Wang Z, Liu D, Varin A, Nicolas V, Courilleau D, Mateo P, Caubere C, Rouet P, Gomez AM, V and ecasteele G, Fischmeister R, Brenner C.

Journal:

Cell Death Dis (2016): e2198

Usage and storage

Intended use

Research Use Only (RUO)

Instrument settings

Fluorescence microplate reader
Excitation	490 nm
Emission	525 nm
Cutoff	515 nm
Recommended plate	Black wall/Clear bottom
Instrument specification(s)	Bottom read mode/Programmable liquid handling

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Page updated on October 13, 2026