資源描述：
《Nuclear Proteins Analysis Tools》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫(kù)。
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.
