Mediomics/Bridge-It® Cyclic AMP (cAMP) all in one Assay Kit, 384-well format/122938

Bridge-It^® CyclicAMP(cAMP)Assays–Selectivity,SensitivityandPerformanceCharacteristics

TheBridge-It^® CyclicAMP(cAMP)assayishighlyspecific.ATP,AMP,andcGMPhaveallbeentestedforselectivityusingthecAMPassay.NoresponsewasdetectedusingtheBridge-It^® cAMPassaywithanyofthesecompoundswithintheconcentrationrangethatwouldbeexpectedtooccurin“reallife”samples(i.e.,millimolarrangeforAMPandATP,andmicromolarrangeforcGMP).Thesensitivityandperformancecharacteristicsofthe Bridge-It^®cAMP designer andtheBridge-It^® allinone assayproductsare presentedinthefollowingTableandFigures.

Pricing: The100-wellkitisaone-timetrialpurchase.Forbulkpricingoptions,please contactus.Foranyotherquestionsorcomments,pleasedonothesitateto contactus.Wearealwayshappytohelp!

ExampleofBridge-It^® cAMP allinone AssayforCellsinSUSPension

TheBridge-It^® cAMP allinone assayallowsforthestimulationandmeasurementofcAMPinadherentmonolayercellsattachedtothewellsofatissueculturemicroplate.Inthefollowingexample,HEK293cellsweretrypsinizedandplatedintothewellsofa96-wellpolystyrenetissueculturemicroplateatadensityof25,000cellsperwellin50µlofculturemedia.Thecellswereallowedtoattachtothebottomofthewellsovernight.Themediawasremovedthenextdayandthewellswerewashedwithabufferedsalinesolution.Thewashbufferwasthenremovedandreplacedwith50µlofKRB-IBMXbuffer.Forstimulation,1µlofforskolinatdifferentconcentrationswasaddedtoeachwellandincubatedandrotatedgentlyfor15minutesat~25ºC.ToquantifycAMPineachwell,theforskolinbuffersolutionwasremoved,100µlofthe allinone AssaySolutionwasaddedtoeachwell,andthemicroplatewasincubatedfor30minutesat~25ºCwithrotation.Thewellcontentswerethentransferredintoablack96-wellmicroplateandthefluorescentsignalwasreadusingthesettingsforfluorescein(~485nmexcitation,~540nmemission).

ExampleofBridge-It®cAMP allinone AssayforAttachedCells

Forinformation ontheassayprinciplepleaseseethe Bridge-ItAssayTechnologyplatform pageordownloadthe protocol.

Thisproductisprotectedbypatentsandpendingpatents.

SelectedCustomerPublicationsusing Mediomics’Bridge-ItcAMPFluorescence AssayKit:

1. Neuronostatininhibitsglucose-stimulatedinsulinsecretionviadirectactiononthepancreaticα-cell.SalvatoriAS,ElrickMM,SamsonWK,CorbettJA,YostenGL. AmJPhysiolEndocrinolMetab.2014Jun1;306(11):E1257-63.

2. 14-Deoxyandrographolidealleviatesethanol-inducedhepatosteatosisthroughstimulationofAMP-activatedproteinkinaseactivityinrats.MandalS,MukhopadhyayS,BandhopadhyayS,SenG,BiswasT. Alcohol.2014Mar;48(2):123-32.

3. Thefractalkine/CX3CR1systemregulatesβcellfunctionandinsulinsecretion.LeeYS,MorinagaH,KimJJ,LagakosW,TaylorS,KeshwaniM,PerkinsG,DongH,KayaliAG,SweetIR,OlefskyJ. Cell.2013Apr11;153(2):413-25.

4. Regulationofforskolin-inducedcAMPproductionbycytochromeP450epoxygenasemetabolitesofarachidonicacidinHEK293cells.AbukhashimM,WiebeGJ,SeubertJM. CellBiolToxicol.2011Oct;27(5):321-32.

5. Netrinspromotedevelopmentalandtherapeuticangiogenesis.WilsonBD,IiM,ParkKW,SuliA,SorensenLK,Larrieu-LahargueF,UrnessLD,SuhW,AsaiJ,KockGA,ThorneT,SilverM,ThomasKR,ChienCB,LosordoDW,LiDY. Science.2006Aug4;313(5787):640-4.

6. Amechanismofcelldeathinvolvinganadenylylcyclase/PKAsignalingpathwayisinducedbytheCry1AbtoxinofBacillusthuringiensis.ZhangX,CandasM,GrikoNB,TaussigR,BullaLAJr. ProcNatlAcadSciUSA.2006Jun27;103(26):9897-902

Thisproductisprotectedby patentsandpendingpatents.

Catalog#: 122939(100measurements-onetimetrialsize),122938(384measurements)

Protocol: 

Bridge-It®cAMPallinoneassaykit,384wellformatProtocol729.92KB

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SDS:

Bridge-It®cAMPallinoneassaykitSDS212.15KB

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