Insituimmunodetectionof5-methylcytosine(5-mC)onsquashedcellsusinganti-methylcytosine

Cross-referencesaretoSchwarzacher&Heslop-Harrison2000.PracticalinsituHybridization.Bios,Oxford.216+xiipp.AvailablefromSigmaChemical

1)Preparechromosomesasforinsituhybridization(Chapter5insitubook)2)Slidesarepre-treatedwithRNaseandpepsin(Protocol8.1insitubookwithoutparaformaldehydetreatment)3)Ethanoldehydratetheslidesandairdry4)BlockslidesinPBST+1%BSAfor20minatRT5)Incubateslidesinmonoclonalanti-5-mC(1:1000dilutioninPBS)for1hat37oCinahumidchamber6)WashinPBST2x5min7)IncubateslidesinBiotinanti-mouseIgG(1:500dilutioninPBST)for1hat37oCinahumidchamber8)WashinPBST2x5min9)IncubateslidesinstreptavidinCy3(1:300dilutioninPBST)for1hat37oCinahumidchamber10)WashinPBST2x5min11)DAPIstainandmount(Protocol9.3insitubook)

Solutions:PBS(seeAppendix1insitubook)PBST:add0.03%(v/v)Tween20tothe1xPBSSeeCastilhoetal.J.CellSci.2000(onPublicationpage)

PatHeslop-Harrison

================  蚂蚁淘在线  ================

免责声明：本文仅代表作者个人观点，与本网无关。其创作性以及文中陈述文字和内容未经本站证实，对本文以及其中全部或者部分内容、文字的真实性、完整性、及时性本站不做任何保证或承诺，请读者仅作参考，并请自行核实相关内容

版权声明：未经蚂蚁淘在线授权不得转载、摘编或利用其他方式使用上述作品。已经经本网授权使用作品的，应该授权范围内使用，并注明“来源：蚂蚁淘在线”。违反上述声明者，本网将追究其相关法律责任。