CA-N226
Name	EliteTMNAD/NADHRatioAssayKit(RedFluorescence)
Description	TheEliteTMFluorimetricNAD/NADHRatioAssayKitprovidesaconvenientmethodforsensitivedetectionofNAD,NADHandtheirratio;thereisnoneedtopurifyNAD/NADHfromsamplemix.IthasverylowbackgroundsinceitisrunintheredvisIBLerangethatsignificantlyreducestheinterferenceresultedfromBIOLOGicalsamples.
Application	Theassaycanbeconvenientlyperformedina96-wellor384-wellmicrotiter-plateformat.ItssignalcanbeeasilyreadbyeitherafluorescencemicroplatereaderatEx/Em=530-570/590-600nm(maximumEx/Em=540/590nm)oranabsorbancemicroplatereaderat~576nm.ThiskitprovidesNADandNADHextractionbuffer,andcelllysisbufferforyourconvenience.IthasbeenfrequentlyusedfordeterminingNAD/NADHfromcelllysates.
Size	250assaysin96-wellplates
Ex/Em	540/590nm
Detection	Fluorescencemicroplatereader

 

Clickherefor DATASHEET

Clickherefor MSDS

 

FrequentlyAskedQuestions 

1.CanweusethisratioassaykittoalsodetectNADorNADHseparately? 

Answer:Yes.YoucouldmeasuretheNADHamountwithComponentD(NADHExtractionSolution),andmeasuretheNADamountwithNADExtractionSolution.Butaccordingtoourexperience,theNADconcentrationsincellsarealothigherthantheNADHconcentrations.WesuggesttomeasuretheTotalNAD+NADHamountandtheNADamount,thentocalculatetheNADHamountbysubtraction,whichgivesmoreaccurateresults.

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.SowhichcurveshouldIusetoquantifymydata? 

Answer:You"llneedtousethecurvesetuponthesameplatewithyoursamples. 

6.Wedon"thavefluorescencemicroplatereader,socanweuseabsorbancemicroplatereader? 

Answer:Yes.Buttheabsorbancereadingshavemuchlowersensitivitycomparedtothefluorescencereadings.Tryusingtheratioof570to610toincreasethesensitivity.

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