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Screen Quest™ Live Cell cAMP Assay Service Pack

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.

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.
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.
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36382100 Tests
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36383200 Tests
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363841000 Tests
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Storage, safety and handling

H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200
Instrument settings

Fluorescence microplate reader
Excitation490 nm
Emission525 nm
Cutoff515 nm
Recommended plateBlack wall/Clear bottom
Instrument specification(s)Bottom read mode/Programmable liquid handling
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Page updated on October 9, 2025