Overview | PrinterFriendlyVersion |
Ex/Em(nm) | 545/576 |
MW | N/A |
CAS# | N/A |
Solvent | Water |
Storage | F/D/L |
Category | EnzymeDetection PeptidasesandProteases |
Related | BiochemicalAssays |
Spectrum | AdvancedSpectrumViewer |
1. Makea5-10mg/mLCasein,TAMRA-conjugatedstocksolutioninPBSbuffer.Unusedstocksolutioncanbedividedintosingleusealiquotsandstoredat-20oC,andavoidexposuretolight.
2. Prepare2XassayworkingsolutionbydilutingtheTAMRA-conjugatedstocksolutioninto50-100mMTrisbuffer(pH7.4)at100-400µg/mL.
Note1:The2XAssayworkingsolutionis designedfordetectingtheactivityofchymotrypsin,trypsin,Thermolysin,proteinaseK,proteaseXIV,andhumanleukocyteelastase.Forotherproteases,pleaserefertoAppendixIfortheappropriateassaybufferformula.
Note2:Theoptimumconcentrationoftheassayworkingsolutionshouldbedeterminedexperimentallyforindividualproteases.
3. Mixequalvolumeofthetrypsinstandardsorsampleswith2XAssayworkingsolution.
4. MonitorthefluorescenceincreaseatEx/Em=540/590nm.
ForkineticreADIng:Immediatelystartmeasuringfluorescenceintensitycontinuouslyandrecorddataevery5minutesfor30minutes.
Forend-pointreading:Incubatethereactionatadesiredtemperaturefor30to60minutes,protectedfromlight.Thenmeasurethefluorescenceintensity.
References&Citations | CitationExplorer |
Liquidtemperaturemeasurementmethodinmicrochannelsbyusingfluorescencepolarization
Authors:KazuyaTatsumi,ChiHsuanHsu,AtsushiSuzuki,KazuyoshiNakabe
Journal:HeatandMassTransfer(2017):1--10
Micro-scaletemperaturemeasurementmethodusingfluorescencepolarization
Authors:KTatsumi,CHHsu,ASuzuki,KNakabe
Journal:(2016):032097
MicroscopicFluidTemperatureMeasurementsUsingFluorescencePolarizationMethod
Authors:KazuyaTatsumi,AkihisaTozaki,KazuyoshiNakabe
Journal:(2011):T10167--T10167