CA-N218 | |
Name | EliteTMNAD/NADHAssayKit(RedFluorescence) |
Description | TheElite™FluorimetricNAD/NADHAssayKitprovidesasensitive,one-stepassaytodetectaslittleas10pico-molesofNAD(H)ina100μlassayvolume(100nM).IthasverylowbackgroundsinceitisrunintheredvisIBLerangethatsignificantlyreducestheinterferenceresultedfromBIOLOGicalsamples. |
Application | Theassaycanbeconvenientlyperformedina96-wellor384-wellmicrotiter-plateformatandreADIlyadaptedtoautomationwithoutaseparationstep.ItssignalcanbeeasilyreadbyeitherafluorescencemicroplatereaderatEx/Em=530-570nm/590-600nm(maximumEx/Em=540/590nm)oranabsorbancemicroplatereaderat~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.