Pglosensor22f camp

Manufactured by Promega

Sourced in United States

The PGloSensor22F-cAMP is a genetically encoded cyclic AMP (cAMP) biosensor. It is designed to measure intracellular cAMP levels in live cells.

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Lab products found in correlation

Glosensor camp assay, by Promega (2 mentions) Co2 independent medium, by Thermo Fisher Scientific (1 mentions) Varioskan lux, by Thermo Fisher Scientific (1 mentions) Flexstation 3 plate reader, by Molecular Devices (1 mentions) Glosensor reagent, by Promega (1 mentions) Hek293ft cells, by Thermo Fisher Scientific (1 mentions) Opti mem, by Thermo Fisher Scientific (1 mentions) X tremegene 9, by Roche (1 mentions) Glosensor camp reagent stock solution, by Promega (1 mentions) Glomax 20 20n luminometer, by Promega (1 mentions)

4 protocols using pglosensor22f camp

Evaluating cAMP Inhibition in Transfected Cells

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cAMP inhibition assays were performed as previously described in HEK293 cells transiently transfected with pGloSensor22F-cAMP (Promega, Madison, WI, USA) and either FLAG-mouse δOR, HA-mouse μOR, or FLAG-mouse κOR.^{40 (link)}

Creed S.M., Gutridge A.M., Argade M.D., Hennessy M.R., Friesen J.B., Pauli G.F., van Rijn R.M, & Riley A.P. (2020). Isolation and Pharmacological Characterization of Six Opioidergic Picralima nitida Alkaloids. Journal of natural products, 84(1), 71-80.

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Measurement of Intracellular cAMP Levels

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HEK293 cells
were transfected with pGloSensor-22F cAMP (Promega Corporation, United
States) and pcDNA3.1-β₂AR using Hilymax. Intracellular
cAMP levels were measured by using GloSensor cAMP assay (Promega Corporation,
United States, #E1290) according to the manufacturer’s protocol.
Briefly, transfected cells were equilibrated with CO₂ independent
medium (Thermo Fisher Scientific, United States, #18045088) containing
10% FBS and 2% GloSensor cAMP reagent. After incubating for 2 h in
room temperature, the cells were exposed to the test samples and subjected
to kinetic measurement of luminescence for the determination of cAMP
levels by a microplate reader (Varioskan Lux, Thermo Fisher Scientific,
United States).

Krama A., Tokura N., Isoda H., Shigemori H, & Miyamae Y. (2022). Cyanidin 3-Glucoside Induces Hepatocyte Growth Factor in Normal Human Dermal Fibroblasts through the Activation of β2-Adrenergic Receptor. ACS Omega, 7(26), 22889-22895.

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Monitoring Cellular cAMP Signaling

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HEK293FT cells (Life Technologies, Grand Island, NY, USA) were transiently transfected with pGloSensor22F-cAMP (Promega, Madison, WI, USA) and pcDNA3.1-FLAG-mDOR in 3:1 DNA mass ratio using XtremeGene9 (Roche, Indianapolis, IN, USA) according to the manufacturer's guidelines. Cells were dislodged 48 hours post-transfection and resuspended in OptiMEM (Fisher Scientific, Pittsburgh, PA, USA) to 3x10 6 cells/mL and seeded in Greiner Bio-One CELLSTAR round-bottom 384-well plates (Fisher Scientific, Pittsburgh, PA, USA) and incubated at 37°C/5% CO 2 for 90 min. Cells were pre-equilibrated with 7.5 μl (4% v/v) GloSensor reagent (Promega, Madison WI, USA) for 90 min at room temperature. Equilibrated cells were stimulated with 4X drug dilution series for 20 min at room temperature prior to the incubation with 31.5 μM forskolin for 20 min at room temperature. A FlexStation3 plate reader (Molecular Devices, Sunnyvale, CA, USA) was used to measure the relative luminescence units (RLU).

Meqbil Y.J., Aguilar J., Blaine A.T., Chen L., Cassell R.J., Pradhan A.A, & van Rijn R.M. (2024). Identification of 1,3,8-Triazaspiro[4.5]Decane-2,4-Dione Derivatives as a Novel δ Opioid Receptor-Selective Agonist Chemotype. The Journal of pharmacology and experimental therapeutics, 389(3).

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Measuring Parapinopsin-Mediated cAMP Levels

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The intracellular cAMP level changes in lamprey parapinopsin-expressing HEK293S cells were measured using the GloSensor cAMP assay (Promega) as previously described [27 (link)]. HEK293S cells (20–30% confluent) in 35-mm tissue culture dishes were transfected with 1.5 μg each of the lamprey parapinopsin and the pGloSensor-22F cAMP (Promega) plasmid using the PEI transfection method, followed by incubation overnight in a culture medium containing 10% FBS. Following the overnight incubation after the supplement of 11-cis retinal to the medium, the medium was replaced with a CO2-independent medium containing 10% FBS and GloSensor cAMP Reagent stock solution (Promega). Luminescence, representing the amount of cAMP, was measured at 25°C using a GloMax 20/20n Luminometer (Promega).

Wada S., Kawano-Yamashita E., Sugihara T., Tamotsu S., Koyanagi M, & Terakita A. (2021). Insights into the evolutionary origin of the pineal color discrimination mechanism from the river lamprey. BMC Biology, 19, 188.

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