CA-N418 | |
Name | EliteTMNADP/NADPHAssayKit(RedFluorescence) |
Description | TheElite™FluorimetricNADP/NADPHAssayKitprovidesasensitive,one-stepassaytodetectaslittleas1picomolesofNADPHina100μlassayvolume(10nM).IthasverylowbackgroundsinceitisrunintheredvisIBLerangethatsignificantlyreducestheinterferenceresultedfromBIOLOGicalsamples.ThereisnoneedtopurifyNADP/NADPHfromsamplemix. |
Application | Theassaycanbeconvenientlyperformedina96-wellor384-wellmicrotiter-plateformat.ItssignalcanbeeasilyreadbyeitherafluorescencemicroplatereaderatEx/Em=540/590nmoranabsorbancemicroplatereaderat~576nm. |
Size | 400assaysin96-wellplates |
Ex/Em | 540/590nm |
Detection | Fluorescencemicroplatereader |
Clickherefor DATASHEET | Clickherefor MSDS |
FrequentlyAskedQuestions
1.AfterIlysethemammaliancells,doIhavetodotheassayrightaway?MayIfreezethesamplesin-80Cfreezeranddotheassayslater?
Answer:Youmayfreezethesamplesat-80Cafterthey"relysed,anddotheassaysaltogether.
2.WhyPBSisusedtodilutetheStandard?Sincethesamplesareinthelysisbuffer,doyouthinkIshouldusethelysisbuffertodilutethem?
Answer:PBSiscommonlyused.Peopleusuallyusethelysisbuffertolysethecells,thenusePBStodilutethesamples.ThepointisthatyoushouldkeepthesameconcentrationofthelysisbufferforthesamplesandtheStandards.
3.Ididacomparison,anditshowedthatthePBSbasedStandardcurvelookedbetterthanthelysisbufferbasedStandardcurve.Whatdoyouthinkofthis?
Answer:ThebubblesinthelysisbuffermaycausetheinconsistencyofthereADIngs.
4.WhenIdoduplicatesofStandards,thereadingvariesalotinthelowrange(0.2uMorlower).Whydoesthathappen?
Answer:Itisnormal.Therearemanyfactorsthatwouldcauseinconsistentreadings,i.e.bubblesinthewells,thevolumes....Sotrydoingmorereplicates.
5.Sometimesforthe2setsofStandardsIsetup,theylookgood(R2=0.99)individuallybutthecurveismessedup(R2=0.95)whenmerged.SowhichcurveshouldIusetoquantitatemydata?
Answer:You"llneedtousethecurvesetuponthesameplatewithyoursamples.
6.Wedon"thavefluorescencemicroplatereader,socanweuseabsorbancemicroplatereader?
Answer:Yes.Buttheabsorbancereadingshavemuchlowersensitivitycomparedtothefluorescencereadings.Tryusingtheratioof570to610toincreasethesensitivity.
7.Wedonothave540nmexcitationfilterbutwehave560nm.Wedon"thavetheabsorbance573nm.Whatistherangewecouldpossibleuse?
Answer:Werecommendtouse560/590,andcutoff570nm;OR560/610,cutoff590nm.