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AAT Bioquest/Biotin-X NTA [Biotin-X nitrilotriacetic acid, potassium salt] *CAS 856661-92-2*/3009/1 mg
Overview | PrinterFriendlyVersion |
Ex/Em(nm) | None/None |
MW | 715.98 |
CAS# | 856661-92-2 |
Solvent | DMSO |
Storage | F/D |
Category | BiotinandItsDerivatives BiotinConjugates |
Related | BiochemicalAssays |
SampleProtocolfordetectionofHis-tagged ProteininPVDF:
1) PrepareyourPVDFblotsasneeded(blocknonspecificbindingsitesandwashtheblot).
2) DissolvetheBiotin-XNTAinDMSOorH2Oatconcentrationof1mg/mL
3) PreparefreshStainingSolution(lessthan30minutesbeforeuse):prepare20mLofStainingSolutionforeach8cm×10cmblotbyadding20μLof10mMNiCl2,20μLof1mg/mLbiotin-XNTA(fromStep2)and1-2μLof1mg/mLstreptavidin–alkalinephosphatase(madeinstep1.2)to20mLofBlockingBuffer.Mixwell.Oncethesolutionismade,itisreadytouserightaway,andremainsstableforatleast30minutes.
4) Incubatetheblotwith20mLofStainingSolution(fromStep3)atroomtemperaturefor30minutes.
5) Washtheblots,andpursuitforchemiluminescencedetection.
SampleProtocolforBiotin-XNTA–ProbedBlotStripping:
1) Ifdesired,theblotcanbestainedusingotherdetectionmethods.
2) Forstainingwithantibodiesorlectins,stripthebiotin-XNTAcomplexofftheblotbyincubatingitin62.5mMTris,0.2%SDS,50mMdithiothreitol(DTT),pH6.8,at50°Cfor40minuteswithgentleagitation.Orincubatedovernightin200mMacetateacidwith40mMEDTA,pH4.8.
3) WashtheblotinWashBufferatroomtemperature2-4timesfor5minuteseachandproceedwithantibodydetection.
References&Citations | CitationExplorer |
Alow-cost,high-performancesystemforfluorescencelateralflowassays
Authors:LindaGLee,EricSNordman,MartinDJohnson,MarkFOldham
Journal:Biosensors(2013):360--373
ImplementationofP22viralcapsidsasnanoplatforms
Authors:SebyungKang,MasakiUchida,AlisonO’Neil,RuiLi,PeterEPrevelige,TrevorDouglas
Journal:Biomacromolecules(2010):2804--2809
Calcium-dependentactivationoftransglutaminase2bynanosecondpulsedelectricfields
Authors:KeikoMorotomi-Yano,Ken-ichiYano
Journal:FEBSOpenBio