醋酸纤维素膜电泳分离血浆蛋白英文版VIP免费

Disciplinesinthelaboratory：1.Mustwearwhitecoatanddon’tbelate.2.Mustkeepquietwhenyouareinthelaboratory.3.Don’ttouchanyexperimentalapparatuswithoutmypermission.4.Anassistantneeded.Collecttheexperimentalreportsandasktwostudentsondutytocleanthelabafterclasseverytime.Theinstructorwillcheckthelaboratoryatlast.Fivescoreswillbelostinyourfinialexaminationifyouwerelateordidnotcleanthelabeachtime.Experimentalreport1.Experimentalprinciple2.Experimentalmaterials(optional)3.Experimentalprocedure4.ResultsandanalysisorResultsandCalculationIfyougetamembrane(picture),pleasepasteitonthelabnotebook.Describethemembrane(picture)andtrytoexplainthepossiblereasonwhyyougettheresult;Ifyougetthemathematicdata,pleasedosomecalculation.5.ConclusionUseoneortwosentencestosummarizeyourexperiment.TheexperimentaltitleSeparationofplasmaproteinsbycelluloseacetatemembraneelectrophoresisElectrophoresisChargedparticlesmovetotheelectrodeofversechargeintheelectricfield.Particles:bio-macromolecule.e.g.DNA,RNAorprotein.Direction:Ifparticlescarrynegativecharge,itwillmovetowardpositiveelectrode.Ifparticlescarrypositivecharge,itwillmovetowardelectrode.negativeCelluloseacetatemembraneAkindofmaterialthathasthenet-likestructure.Thespecialstructurecanretardtheproteinsbythemolecularweight.Proteinlarger,movesslowerintheelectricfield.ProteinsintheplasmaThousandsofproteinsintheplasma.Themainproteinsarealbumin,α1-globulin,α2-globulin,β-globulin,fibrinogenandγ-globulin.Theseproteinscanbeseparatedbycelluloseacetatemembraneelectrophoresis.1.ExperimentalprincipleTheisoelectricpoint(pI),isthepHatwhichaparticularmoleculecarriesnonetelectricalchargeintheenvironmentalbuffer.1.ExperimentalprincipleDifferentproteinsinplasmahavedifferentisoelectricpoints(pI),sotheyhavedifferentquantityofelectricalchargeintheelectrophoreticbuffer(pH8.6).Moreover,proteinsalsohavedifferentmolecularweightsandshapes.Underthesameconditionofelectricfield,theywillmoveatthedifferentspeeds,whichiscalledelectrophoreticmobility.Afteraperiodoftime,proteinsinplasmacanbeseparatedbyelectrophoresis.Whataffectselectrophoreticmobility?Externalfactors:electric-fieldstrength,pHvalueinthebuffer,ionicstrength,etc.Internalfactors:quantityofelectriccharge、molecularweightandshapes.2.Materials(optional)Celluloseacetatemembrane(2×8cm)PowersupplyCoverslip(loadingsample)filterpaperslideglasstweezerHumanplasmabarbitalbuffer(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.Saltbridgeofpositiveelect...