![AAT Bioquest/FDP[荧光素二磷酸四铵盐]*CAS 217305-49-2*/11600/5 mg](https://www.ebiomall.cn/images/AAT201712/chemical-structure-for-fdp-fluorescein-diphosphate-tetraammonium-salt-cas-217305-49-2.png)
| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 490/514 |
| MW | 560.39 |
| CAS# | 217305-49-2 |
| 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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