![醋酸纤维素膜电泳分离血浆蛋白(英文版)_第1页](http://file4.renrendoc.com/view11/M00/0C/28/wKhkGWVsQ-KACBYgAAM0gs3SyBE156.jpg)
![醋酸纤维素膜电泳分离血浆蛋白(英文版)_第2页](http://file4.renrendoc.com/view11/M00/0C/28/wKhkGWVsQ-KACBYgAAM0gs3SyBE1562.jpg)
![醋酸纤维素膜电泳分离血浆蛋白(英文版)_第3页](http://file4.renrendoc.com/view11/M00/0C/28/wKhkGWVsQ-KACBYgAAM0gs3SyBE1563.jpg)
![醋酸纤维素膜电泳分离血浆蛋白(英文版)_第4页](http://file4.renrendoc.com/view11/M00/0C/28/wKhkGWVsQ-KACBYgAAM0gs3SyBE1564.jpg)
![醋酸纤维素膜电泳分离血浆蛋白(英文版)_第5页](http://file4.renrendoc.com/view11/M00/0C/28/wKhkGWVsQ-KACBYgAAM0gs3SyBE1565.jpg)
版权说明:本文档由用户提供并上传,收益归属内容提供方,若内容存在侵权,请进行举报或认领
文档简介
Disciplinesinthelaboratory:Mustwearwhitecoatanddon’tbelate.Mustkeepquietwhenyouareinthelaboratory.Don’ttouchanyexperimentalapparatuswithoutmypermission.Anassistantneeded.Collecttheexperimentalreportsandasktwostudentsondutytocleanthelabafterclasseverytime.Theinstructorwillcheckthelaboratoryatlast.Fivescoreswillbelostinyourfinialexaminationifyouwerelateordidnotcleanthelabeachtime.Experimentalreport1.Experimentalprinciple2.Experimentalmaterials(optional)3.Experimentalprocedure4.Resultsandanalysis
orResultsandCalculationIfyougetamembrane(picture),pleasepasteitonthelabnotebook.Describethemembrane(picture)andtrytoexplainthepossiblereasonwhyyougettheresult;Ifyougetthemathematicdata,pleasedosomecalculation.
5.ConclusionUseoneortwosentencestosummarizeyourexperiment.
TheexperimentaltitleSeparationofplasma
proteins
bycelluloseacetatemembraneelectrophoresisElectrophoresisChargedparticlesmovetotheelectrodeofversechargeintheelectricfield.Particles:bio-macromolecule.e.g.DNA,RNAorprotein.Direction:Ifparticlescarrynegativecharge,itwillmovetowardpositive
electrode.Ifparticlescarrypositivecharge,itwillmovetowardelectrode.negativeCelluloseacetatemembrane
Akindofmaterialthathasthenet-likestructure.Thespecialstructurecanretardtheproteinsbythemolecularweight.Proteinlarger,movesslowerintheelectricfield.Proteins
intheplasma
Thousandsofproteinsintheplasma.Themainproteinsarealbumin,
α1-globulin,α2-globulin,β-globulin,fibrinogenand
γ-globulin.Theseproteinscanbeseparatedbycelluloseacetatemembraneelectrophoresis.
1.Experimental
principleThe
isoelectricpoint
(pI),isthe
pH
atwhichaparticular
molecule
carriesnonet
electricalcharge
inthe
environmentalbuffer.
1.Experimental
principle Differentproteinsinplasmahavedifferentisoelectric
points(pI),sotheyhavedifferentquantity
of
electrical
chargeintheelectrophoreticbuffer(pH8.6).Moreover,proteinsalsohavedifferentmolecularweightsandshapes.Underthesameconditionofelectricfield,theywillmoveatthedifferentspeeds,whichiscalledelectrophoreticmobility.Afteraperiodoftime,proteinsinplasmacanbeseparatedbyelectrophoresis.Whataffectselectrophoreticmobility?
External
factors:electric-fieldstrength,pH
valueinthebuffer,ionic
strength,etc.Internalfactors:
quantity
of
electric
charge、molecularweightandshapes.2.Materials(optional)Celluloseacetatemembrane(2×8cm)PowersupplyCoverslip(loadingsample)filterpaperslideglass
tweezerHumanplasmabarbitalbuffer(pH8.6)stainingsolution(Amidoblack10B)Washbuffer3.Experimentalprocedure(1).Soakofmembrane:
Makealoadingsamplelinewithapencilonthenon-reflectivesurface.Thelineshouldbeperpendiculartothelengthofmembraneandatabout1.5cmdistancefromthewidthofmembrane.Then,putthemembraneintotheelectrophoreticbufferandsoakitabout30minutes.
(2).Loadingsample: Pickupamembranewithatweezerandabsorbabundantbufferwithfilterpaper.Useacoversliptodiptheplasmaandloadthesampleontheloadingsamplelineofmembraneonlyonce.(Ifyourepeatloadingsamples,theproteinbandsmaybecurve.)(3).Proteinsseparationbyelectrophoresis:
Putthemembraneoverandclosetosaltbridges.Keepthemembranesurfaceofloadingsamplefacingdown.Letthemembranebeperpendiculartothesaltbridgesandkeeptheendofmembraneneartoloadinglinetightlyatsaltbridgeofnegativeelectrode.
Theprofileofelectrophoretictank1.Saltbridgeofnegativeelectrode;2.Electrophoreticbuffer3.celluloseacetatemembrane4.SaltbridgeofpositiveelectrodeDon’tlettheloadinglinetouchwithsaltbridges. Covertheelectrophoretictankwiththelidandsetthevoltageat120volts,runningabout30~45minutes.(3).Proteinsseparationbyelectrophoresis:(4).Dyingthemembrane: Pickupthemembranewithtweezersanddyetheproteinswithstainingsolution(Amidoblack10B)forabout1~3minutes.(5).Washingthemembrane: Washthestainingmembranewiththewashingbuffer
threetimesintheglassvat.Afterwashing,thedarkblueproteinbandscanbevisualizedonthemembrane.Glassvat#1StainingvatGlassvat#3Glassvat#24.Resultsandanalysis(-)(+)Whythestainingcolorofproteinsaredifferent?Whydoessomeproteinmovefaster?Andwhydoessomeproteinmoveslower?Becausethequantityofalbuministhemostamongtheseproteins,thecolorofalbuministhemostdark.Onthecontrary,thequantitiesofα1-globulinandα2-globulinarerelativelyless,sotheircolorareweakerthanotherproteinbands.
Albuminmovesfastestbecauseitsmolecularweightisthesmallestamongtheseproteins.Onthecontrary,γ-globulinmovesrelativelyslowerbecausethemolecularweightarelargerthanotherproteins.
4.ResultsandanalysisThesignalsofproteinbandsarenotverystrong.Itispossiblethattheloadingsampleislessorstainingtimeisnotenough.Theproteinbandsarenotconsecutivebecausetheloadingsampleisuneven.
4.ResultsandanalysisTheproteinbandslookstooclosesothatsomeproteinsarenotseparatedbycelluloseacetatemembraneelectrophore
温馨提示
- 1. 本站所有资源如无特殊说明,都需要本地电脑安装OFFICE2007和PDF阅读器。图纸软件为CAD,CAXA,PROE,UG,SolidWorks等.压缩文件请下载最新的WinRAR软件解压。
- 2. 本站的文档不包含任何第三方提供的附件图纸等,如果需要附件,请联系上传者。文件的所有权益归上传用户所有。
- 3. 本站RAR压缩包中若带图纸,网页内容里面会有图纸预览,若没有图纸预览就没有图纸。
- 4. 未经权益所有人同意不得将文件中的内容挪作商业或盈利用途。
- 5. 人人文库网仅提供信息存储空间,仅对用户上传内容的表现方式做保护处理,对用户上传分享的文档内容本身不做任何修改或编辑,并不能对任何下载内容负责。
- 6. 下载文件中如有侵权或不适当内容,请与我们联系,我们立即纠正。
- 7. 本站不保证下载资源的准确性、安全性和完整性, 同时也不承担用户因使用这些下载资源对自己和他人造成任何形式的伤害或损失。
最新文档
- Unit 1 The mass media Reading 教学设计-2024-2025学年高中英语译林版(2019)选择性必修第二册
- 简历小升初小升初简历
- XX单位中秋节廉政提醒讲话稿
- 2024秋季巴黎奥运会开学第一课主题班会课件
- FMEA工作坊实操课件
- 株洲市第五中学招聘笔试真题2023
- 延安延川县银龄讲学计划笔试真题2023
- 七年级数学上册数学 2.3 数轴(七大题型)(解析版)
- 安康白河县纪委监委选调笔试真题2023
- 结构力学优化算法:灵敏度分析:结构动力学优化与灵敏度分析
- 风力发电机组风机叶片的维护与维修方法及要求
- 无人机航拍业务拓展策划书
- 中国共享经济市场研究报告
- 纯中药面膜培训培训课件
- 微波技术基础课件
- 渐冻症护理查房
- 2023年上海市16区高考语文二模卷【文言文阅读题】汇集练附答案解析
- U8API开发手册(C#版)文档
- 道德与法制学情分析报告
- 缩短门诊等候时间品管圈课件
- 2023年11月贵州省遵义市正安县招考聘用合同制辅警35人笔试历年高频考点-难、易错点荟萃附答案带详解
评论
0/150
提交评论