| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | None/None |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | CellBIOLOGy CellMetabolism |
| Related | RedoxEnzymes BiochemicalAssays |
- Thawallthecomponentsatroomtemperaturebeforeuse.
- Add0.5mLH2OtooneNADPHvial(ComponentD)tomake40XNADPHsolution.
Note:OnevialofNADPHisformaking10mLNADPRegeneratingSolution.
- Add100µLH2OtoNADPRegeneratingEnzymeMix(ComponentC)vialtomake400Xstocksolution.
Note:AliquotandstoreunusedComponentCat<-20oC,keepfromlightandavoidrepeatedfreeze-thawcycles.
4. Prepare2XNADPRegeneratingSolutionaccordingtothefollowingtables,protectedfromlight.
Table1NADPRegeneratingSolutionforone96-wellplate
Components | Volume |
NADPRegeneratingBufferI | 5mL |
NADPRegeneratingBufferII | 5mL |
NADPH | 0.5mL |
NADPRegeneratingEnzymeMix | 50μL |
Totalvolume | 10.55mL |
Note1:ActivatedNADPregeneratingsolutionisnotstable,andshouldbepreparedfreshlybeforeuse.
Note2:10mLofNADPRegeneratingSolutionisenoughfor1plate.AliquotandstoreunusedComponentA,B,C,Dat<-20oC,keepfromlightandavoidrepeatedfreeze-thawcycles.
5. Addequalvolumeof2XNADPRegeneratingSolution(fromStep4)intothedesiredassaysystem.
| References&Citations |  CitationExplorer |
exo-Brevicominbiosynthesisinthefatbodyofthemountainpinebeetle,Dendroctonusponderosae
Authors:MinminSong,AndrewGorzalski,TrangTNguyen,XibeiLiu,ChristopherJeffrey,GaryJBlomquist,ClausTittiger
Journal:Journalofchemicalecology(2014):181--189
exo-BrevicominbiosyntheticpathwayenzymesfromtheMountainPineBeetle,Dendroctonusponderosae
Authors:MinminSong,PatrickDelaplain,TrangTNguyen,XibeiLiu,LeahWickenberg,ChristopherJeffrey,GaryJBlomquist,ClausTittiger
Journal:Insectbiochemistryandmolecularbiology(2014):73--80
FunctionalcharacterizationofmyrcenehydroxylasesfromtwogeographicallydistinctIpspinipopulations
Authors:MinminSong,AmyCKim,AndrewJGorzalski,MarinaMacLean,SharonYoung,MatthewDGinzel,GaryJBlomquist,ClausTittiger
Journal:Insectbiochemistryandmolecularbiology(2013):336--343
Aninsect-specificP450oxidativedecarbonylaseforcuticularhydrocarbonbiosynthesis
Authors:YueQiu,ClausTittiger,ClaudeWicker-Thomas,GaelleLeGoff,SharonYoung,EricWajnberg,ThierryFricaux,NathalieTaquet,GaryJBlomquist,RenéFeyereisen
Journal:ProceedingsoftheNationalAcademyofSciences(2012):14858--14863
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