Nuclear Proteins Analysis Tools

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1、ThermoScientificPierceNuclearProteinsAnalysisToolsTranscriptionFactorResearchMadeEasyEMSA(GelShiftAssays)ChIP(Chromatin-Immunoprecipitation)High-ThroughputTranscriptionFactorAssaysCellomicsHCSReagentKitsNonradioactiveEMSAThetraditionallabelforEMSAis32P.Wedev

2、elopedtheThermoScientificLightShiftChemiluminescentEMSAKittoprovidethesamesensitivityandspeedas32Passays,butwithoutthehazardsofradioactivity.Thiskithasbeenwidelyreferencedinrecentyears1-8.Theset-upofanEMSA-experimentcanbequiteachallenge.Weofferproductsthatwi

3、llguideevenresearchers,whoarenewtothistechnique,successfullyfromstarttofinish.1.Step:TheSamplePreparationForbestsensitivityandspecificity,mostresearcherswillusenuclear100CytoplasmextractsasstartingmaterialinexperimentslikeEMSA.Thepreparation90Nucleusofnuclea

4、rextractscanbetime-consumingandcompromisetheintegrity80ofmanyfragilenuclearproteins.ThermoScientificNE-PERNuclearandCytoplasmicExtractionKit(#78833)1,3-8enablesyoutostepwise70generatebothfunctionalcytoplasmicandnuclearextractsquicklyand60conveniently.Ourpre-

5、aliquotedThermoScientificHaltProtease504,6,7%SignalInhibitorCocktail(#78425)willprotecttheintegrityofyour40extracts.Becausemanytranscriptionfactorsarephosphoproteins,theadditionofourpre-aliquotedThermoScientificHaltPhosphatase30InhibitorCocktail(#78428)isrec

6、ommendedaswell.20100Oct-1β-GalHsp90•Fast–obtainnuclearandcytoplasmicfractionsinlessthantwohoursNuclearCytoplasmicCytoplasmic•Extractscanbemadefromcellcultures2-8ortissues1LocalizedProteinLocalizedProteinLocalizedProtein•NuclearextractcanbedirectlyusedintheLi

7、ghtShiftKit(when10%ofthetotalWesternBlotActivityAssayWesternBlotreactionvolumeisnotexceeded)ormaybedesaltedwithourThermoScientificSlide-A-LyzerMINIorZebaUnits3Figure1.TheThermoScientificNE-PERReagent’sstepwiseextractionprocessresultsin•Highproteinyieldandmin

8、imal(<10%)cross-contaminationbetweenfractionsminimalcross-contaminationbetweencytoplasmicandnuclearfractions.Oct-1andHsp90(Figure1)determinedbyWesternblotanalysis.β-Galdeterminedbyactivityassay.