Formulation | 50%glycerol/water(v/v) |
Storage | -20°C |
Purity | >95%bySDS-PAGE |
ActivityDetermination | FactorXclottingassayorchromogenicassay |
ShelfLife(properlystored) | 12months |
TheparticipationoftheserineproteasefactorXaintheprothrombinasecomplexisillustrated.MembraneboundfactorXabindstomembraneboundfactorVatoformtheprothrombinasecomplex.Thiscomplexeffectivelyconvertsthezymogenprothrombin(II)totheactiveserineproteasethrombin(IIa)byproteolyticremovalofthefragment1.2(F1.2)portionofprothrombin.
SampleGelInformation:
Gel | Novex4-12%Bis-Tris |
---|---|
Load | HumanFactorXa,1µgperlane |
Buffer | MOPS |
Standard | SeeBluePlus2;Myosin(191kDa),PhosphorylaseB(97kDa),BSA(64kDa),GlutamicDehydrogenase(51kDa),AlcoholDehydrogenase(39kDa),CarbonicAnhydrase(28kDa),MyoglobinRed(19kDa),Lysozyme(14kDa) |
SpecialNotes | Heavychainisadoubletduetothepresenceofupto50%betaform.Theconversionofalpha-Xatobeta-Xaoccursbyautocleavageofalpha-Xabyalpha-XaresultinginthelossofaCOOH-terminalpeptide. |
Overview:
Activationofthezymogen,factorX,byeithertheintrinsicorextrinsicfactorXasecomplexesproducestheactiveserineproteasefactorXa(1,2).TheactivationoffactorXrequiresproteolyticcleavageoftheheavychain,resultinginthereleaseofanactivationglycopeptide.TheheavychainregioninfactorXacontainstheserineproteasecatalyticdomain,whilethelightchain,asinthezymogen,containsthemembranebindingdomain.
FactorXaparticipatesintheprothrombinasecomplex,whichcatalyzestherapidconversionofprothrombintothrombin.ProthrombinaseisanenzymecomplexcomposedoffactorXa(enzyme)andfactorVa(cofactor)assembledonacellularsurfaceinthepresenceofcalciumions.AlthoughfactorXacanindependentlycatalyzetheactivationofprothrombin,therateatwhichthisreactionoccursisincreasednearly300,000-foldwithcompleteassemblyoftheprothrombinasecomplex.TheclottingactivityoffactorXainvivoisterminatedbyeitherinactivationofthecofactor,factorVa,orbydirectinhibitionoffactorXabyinhibitors,suchasATIII,afterdisassemblyoftheprothrombinasecomplex.
Inrecentyears,molecularBIOLOGistshaveutilizedfactorXaforsitespecificcleavageoffusionproteinsexpressedinbacteria(9-12).AfactorXa-sensitivesiteisincorporatedbetweentherecombinantproteinofinterestandpeptidesorproteinswhichfacilitatepurificationand/orexpression.ThetargetproteinisreleasedfromtheexpressedhybridbycleavagewithfactorXa.ThefactorXacanthenbeeasilyremovedbyaffinitychromatography.
FactorXaispreparedbyactivatingpurifiedfactorXwiththefactorXactivatorisolatedfromRussell"svipervenom.FactorXaispurifiedfromtheactivationmixturebychromatographyoverbenzamidine-Sepharosefollowedbygelfiltration(1,3).SeveralmodifiedformsoffactorXaarealsoavailableincluding:A)active-siteblockedfactorXacontainingeitherthetripeptidechloromethylketoneinhibitorEGRck,orthefluorescentinhibitorDansyl-EGRck;andB)humanGla-domainlessβ-factorXa.Theenzymeissuppliedin50%(vol/vol)glycerol/H2Oandshouldbestoredat-20°C.PurityisdeterminedbySDS-PAGEanalysisandactivityismeasuredinafactorXaclottingassayand/orchromogenicsubstrateassay.
InadditiontoitsbroadapplicationincoagulationresearchfactorXacanbeusedforsitespecificcleavageoffusionproteins.AfactorXasensitivesiteisincorporatedbetweentherecombinantproteinofinterestandpeptidesorproteinswhichfacilitatepurificationand/orexpression.ThetargetproteinisreleasedfromtheexpressedhybridbycleavagewithfactorXa.FactorXacanthenbeeasilyremovedbyaffinitychromatography.LottolotconsistencyensuresreproducIBLeresultseverytime.Forexperimentsinvolvingcellcultures,pleasecontactustodiscusscustom,lowendotoxinlotsdesignatedforcellcultureuse.
Properties:
Localization | Plasma | ||||||||
---|---|---|---|---|---|---|---|---|---|
Modeofaction | Enzymecomponentoftheprothrombinasecomplex | ||||||||
Molecularweight | 46,000(human)(4) 45,300(bovine)(5) | ||||||||
Extinctioncoefficient |
| ||||||||
SpecificActivity | approximately1000units/mg | ||||||||
Structure | twosubunits,Mr=16,200and29,000(human)(6),Mr=16,500and28,800(bovine)(5),NH2-terminalgladomain,twoEGFdomains | ||||||||
Percentcarbohydrate | 3.0%(human)(8) 2.1%(bovine)(8) | ||||||||
Post-translationalmodifications | elevenglaresidues, oneβ-hydroxyaspartate |
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风疹病毒抗体IgM阳性才需要治疗。
2-IgM也是一种特异性感染指标,但在人体内持续时间不长(有时是30天更短),往往是传染病的近期感染指标。呈阴性是指:你近期没有感染过结核杆菌。
1,IgG抗体是抗体中分子量最小的一种,可通过胎盘输给胎儿,保护了婴儿最初六个月内免受感染。该抗体产生晚,维持时间长,消失慢,浓度高。血中检测到可作为远期感染指标。
2,IgM抗体是抗体中分子量最大的一种,不能通过胎盘输给胎儿。抗体产生最早,一经感染,快速产生,在感染初期抗感染起作用。但维持时间短,消失快。
3,灵长类动物主要有四种免疫球蛋白IgG、IgM、IgA、和IgE。
所有用于检测抗原的免疫学方法经适当改良后,均可用于抗体的检测,如IFA、ELISA、RIA、LA......由于抗原方法的敏感性提高和PCR技术的应用,使得HSV抗体在HSV感染个体中的不均一性和不稳定性影响了这类指标在临床诊断中的意义,但作为一种感染有关指标,在一定的范围和情况下,仍有必要进行检测和深入研究.目前HSV特异性抗体的检测,主要有IgG、IgM和IgA三种。
IgM是免疫球蛋白M(Immunoglobulin M,IgM)的缩写。根据结构的不同将免疫球蛋白分为五种,IgM是人的免疫球蛋白之一,其他还有lgA、lgG、IgD和lgE。