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						AAT Bioquest/MUP,二钠盐[4-甲基伞形磷酸二钠盐]*CAS 22919-26-2*/11612/10 g
					
								| Overview | PrinterFriendlyVersion  | 
| Ex/Em(nm) | 360/449 | 
| MW | 300.11 | 
| CAS# | 22919-26-2 | 
| Solvent | Water | 
| Storage | F/D/L | 
| Category | EnzymeDetection Phosphatases  | 
| Related | ProteinPhosphatase BiochemicalAssays  | 
| Spectrum | AdvancedSpectrumViewer | 
1.Prepareworkingsolution:
1.1 Preparea2to10mMstocksolutioninanappreciatesolvent(seeTable).Thestocksolutionshouldbeusedpromptly.Anyunusedsolutionneedtobealiquotedandfrozenat<-20oC.
Note1:DonotuseDMSO,ETOHorMETHtomakestocksolutionofSunRed™Phosphate(cat#11629)sinceitsignificantlyincreasesassaybackground.
Note2:Avoidrepeatedfreeze-thawcycles,andprotectfromlight.
1.2 Preparea2XPhosphateworkingsolution: Onthedayoftheexperiment,eitherdissolvethesubstrateinanappreciatesolventorthawanaliquotofthestocksolution(fromStep1.1)atroomtemperature.Preparea2Xworkingsolutionof10to50µMin100mMTrisbufferorbufferofyourchoice,pH8to9(notphosphatebuffer).
2.Runphosphataseassayinsupernatants:
2.1 Add50µLof2XPhosphateworkingsolution(fromStep1.2)intoeachwellofthephosphatasestandard,blankcontrol,andtestsamplestomakethetotalphosphataseassayvolumeof100µL/well.
Note:Fora384-wellplate,add25µLofsampleand25µLof2XPhosphateworkingsolutionintoeachwell.
2.2 Incubatethereactionfor30to120minutesatthedesiredtemperature,protectedfromlight.
2.3 Monitorthefluorescenceincreaseatanappropriatefiltersets(Seetable)withafluorescenceplatereader.
2.4 Thefluorescenceinblankwells(withtheassaybufferonly)isusedasacontrol,andissubtractedfromthevaluesforthosewellswiththephosphatasereactions.
| References&Citations | PrinterFriendlyVersion  | 
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