| Formulation | 50%glycerol/water(v/v) |
| Storage | -20°C |
| Purity | >95%bySDS-PAGE |
| ActivityDetermination | Clottingassay |
| ShelfLife(properlystored) | 12months |
ThedomainstructureofhumanfactorVIIisrepresented,where:GLA=regioncontainingγ-carboxyglutamicacidresidues,EGF=regioncontainingsequenceshomologoustohumanepidermalgrowthfactor,CATALYTICDOMAIN=regioncontainingtheserineproteasecatalytictriad.ThesiteatwhichfactorXacleavesfactorVIItoformfactorVIIaisindicatedwithanarrow.
SampleGelInformation:

| Gel | Novex4-12%Bis-Tris |
|---|---|
| Load | HumanFactorVII,1µgperlane |
| Buffer | MOPS |
| Standard | SeeBluePlus2;Myosin(191kDa),PhosphorylaseB(97kDa),BSA(64kDa),GlutamicDehydrogenase(51kDa),AlcoholDehydrogenase(39kDa),CarbonicAnhydrase(28kDa),MyoglobinRed(19kDa),Lysozyme(14kDa) |
Overview:
HumanfactorVIIisasinglechain,vitaminK-dependent,plasmaglycoproteinwhichissynthesizedintheliver(1-3).Priortosecretionintotheblood,posttranslationalmodificationbyavitaminK-dependentcarboxylaseproducesten-carboxyglutamicacid(gla)residueslocatedintheNH2-terminalportionofthemolecule,whichfacilitatecellmembranebinding.FactorVIIisproteolyticallyactivatedtotheserineprotease,factorVIIa,duringcoagulation.FactorVIIcanbeactivatedbythrombin,factorIXa,factorXaorfactorXIIa.TheactivationresultsincleavageofthesinglechainmoleculeontheCOOH-terminalsideofarginine-152,toproduceanNH2-terminalderivedlightchain(Mr=20,000)andaCOOH-terminalderivedheavychain(Mr=30,000)whichremaincovalentlyassociatedbyasingledisulfidebond.Thelightchainregioncontainsthegladomain,aswellastwogrowthfactordomainswhicharehomologoustohumanepidermalgrowthfactor(EGF).Asingleβ-hydroxyasparticacididentifiedinfactorVIIisalsolocatedinthelightchainregion.TheheavychainregionoffactorVIIacontainsthecatalyticdomain.FactorVIIaandthecofactor,tissuefactor,maycombineonnegativelychargedcellsurfacesinacalciumdependentmannertoformtheextrinsicfactorXaseenzymecomplex.ThisenzymecomplexcatalyzestheconversionofbothfactorIXtofactorIXaandfactorXtofactorXa.TheCDNAforfactorVIIhasbeenisolatedandthenucleotidesequencedetermined(4).FactorVIIsharesextensivesequencehomologywithotherserineproteasesincludingfactorIX,factorXandproteinC.
HumanfactorVIIispurifiedusingacombinationofconventionaltechniques(2)andimmunoaffinitychromatography(5).Thepurifiedproteinissuppliedin50%(vol/vol)glycerol/H2Oandshouldbestoredat-20oC.PurityisdeterminedbySDS-PAGEanalysisandactivityismeasuredinafactorVIIclottingassay.
Properties:
| Localization | Plasma | |||||
|---|---|---|---|---|---|---|
| Plasmaconcentration | 0.5µg/ml(2)-(baseduponactivitymeasurements) | |||||
| Modeofaction | Zymogen;precursortotheserineproteasefactorVIIa | |||||
| Molecularweight | 50,000(2) | |||||
| Extinctioncoefficient |
| |||||
| Isoelectricpoint | 4.8-5.1(6)-(baseduponanalysisofbovinefactorVII) | |||||
| Structure | singlechain,NH2-terminalgla-domain,twoEGFdomains | |||||
| Percentcarbohydrate | 13%(7)-(baseduponanalysisofbovinefactorVII) | |||||
| Post-translationalmodifications | oneβ-hydroxyaspartate(8),tenglaresidues(9) |
ebiomall.com
>
>
>
>
>
>
>
>
>
>
>
>
本人目前需从电鳗放电器官中提取乙酰胆碱受体蛋白,提纯需要的步骤和电鳗器官都可以提供,有没有提供外包提纯服务的公司或者实验室。
NaturalProteinStopsDeadlyHumanBrainCancerInMice
ScientistsfromJohnsHopkinsandfromtheUniversityofMilanhaveeffectivelyproventhattheycaninhibitlethalhumanbraincancersinmiceusingaproteinthatselectivelyinducespositivechangesintheactivityofcellsthatbehavelikecancerstemcells.ThereportispublishedinNature.
Themostcommontypeofbraincancer-glioblastoma-ismarkedbythepresenceofthesestem-cell-likebraincells,which,insteadoftriggeringthereplacementofdamagedcells,formcancertissue.Stemcells,unlikeallothercellsinthebody,arecapableofformingalmostanykindofcellwhentheright"signals"triggertheirdevelopment.
Fortheirtreatmentexperiment,theresearchersreliedonaclassofproteins,bonemorphogenicproteins,thatcauseneuralstem-cell-likeclusterstolosetheirstemcellproperties,whichinturnstopstheirABIlitytodivide.
Firsttheypretreatedhumanglioblastomacellswithbonemorphogenicprotein4(BMP4),theninjectedthesetreatedcellsintomousebrains.Inmiceinjectedwithcellsthatwerenotpretreated,large,invasivecancersgrew.InthemicewithBMP4-treatedcells,nocancersgrewatall.Threetofourmonthsafterinjection,allmicethatgotuntreatedcellsdied,andnearlyallmicewithBMP4-treatedcellswerealive.
Next,thescientistsdeliveredslow-releaseBMP4-containing"beads"directlyintomousebrainswithimplantedglioblastomacells.Micethatgotemptybeadsdevelopedlargemalignanttumorsanddied.MicewithBMP4beadssurvivedmuchlonger,and80percentsurvivedfourmonthsaftercancercellimplants.
"OurideaistotreatpatientswithBMP4orsomethinglikeitrightaftersurgerytoremoveglioblastomainhopesofpreventingtheregrowthofthecancerandimprovingsurvivaltime,"saysAlessandroOlivi,M.D.,directoroftheDivisionofNeurosurgicalOncologyatHopkinsandacontributortothestudy.
OlivisaysclinicalstudiesusingBMP4couldbeginwithinayearand,ifsuccessful,drugtherapiescouldbeavailabletothepublicwithinthreetofouryears.
"ThiswasproofoftheideathatBMPscouldstopglioblastomabydepletingthestem-cell-likepopulationthatfeedsit,"saysHenryBrem,M.D.,chairmanoftheDepartmentofNeurosurgeryatHopkinsandacollaboratorinthestudy."Thisopensexcitingdoorstofutureresearchintotreatmentsandtherapiesforsuchadevastatingdisease."

