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Crystallization Trials
蚂蚁淘
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Materials Procedure Notes TakenfrommethodsbyJ.JancarikandS.H.Kim IfyouusemethodsfromtheKinesinsite,weaskthatyoucitetheKinesinHomePageandauthors,ortheappropriatesourcepublicationinyourwork.
================ 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。MiliporefiltertypeHA0.45micron Cultureplates(Linbromodel76-033-05) ProteininDDWorHEPESbuffer(10mg/mL) Vacuumgrease Syringewith18Gneedle 1. Milliporefilterallsolutionstobeused. 2. Dialyzeproteinfor24hoursagainstwater,ifpossIBLe.Otherwise,includeminimumrequirementsforkeepingtheproteinsoluble.IfbufferisnecessaryuseHEPESbuffer. 3. Fortheinitialscreeningusethebuffersandprecipitantsrecommendedinthefastcrystalscreen(Jancarik,J.&Kim,S.H.1991J.Appl.Cryst.24,409),asinstructed,andthehangingdropmethodwiththeLinbrotissuecultureplatesthathave24wells,1.7cmdiameter,1.6cmdeep.Seediagram. 4. Useaproteinsolutionwithastartingconcentrationof10mg/mL.Thisconcentrationmayhavetobeadjusteddependingonthenumberofthedropsthatprecipitate.Theconcentrationisaboutcorrectwhenone-thirdtoone-halfofthedropsdevelopprecipitatewithinoneweek. 5. Deposit0.7mLofreservoirsolutionintothewell.Combine2microlitersoftheproteinsolutionand2microlitersofthereservoirsolutiontomakeuptheproteindroplet. 6. Incubateoneatroomtemperatureandoneatcold(4-10°C)temperature. 1. CrstallizationscreeningkitsandothersuppliesareavailablefromHamptonresearch.
