Description:
Technozym®uPACombiActibind®ELISA isanassay kit forthecombinedquantitativedeterminationofconcentrationandactivityofu-PAinhumanplasma.u-PAActibind® assay isbasedonacatchingantibodywhichdoesnotinterferewiththefunctionalactivityofu-PA.Followingthebindingofu-PAinthesamplebytheantibody,functionalactivityofboundu-PAisdeterminedusingGlu-Plasminogenandalowmolecularweightplasminsubstrate.Aftermeasuringthefunctionalactivity,thisdeterminationsystemiswashedawayandboundu-PAantigenisdetectedusingaperoxidase-labeledmonoclonalantiu-PAantibodywhichrecognizesbothfreeu-PAandu-PAinhibitorcomplexes.
Specificity:
TheTechnozym®Actibind®u-PAtestmeasuresu-PAfunctionalactivity,noncomplexedu-PAantigenandu-PA-inhibitorcomplexes.Itisnotaffectedbythepresenceofotherplasminogenactivators.Thetestsystemmeasuresu-PAantigeninarangefrom0-25ng/mlanduPAactivityfrom0-2.5IU/ml.Theinter-andintra-assayvariationsarelessthan10%and5%,respectively.
KitComposition:
- Plate+PlateCover
12x8wellplasticmicrotitrestripsprecoatedwithamonoclonalanti-u-PAcoatingantibodyinbicarbonatebuffer,1%bovineserumalbumin(BSA),(TC-CodeGU). - Standard
1xlyophilizedurokinase,calibratedagainstNIBSC87/594(TC-CodeAQ). - POX-Antibody
1xconjugatedpolyclonalantiu-PAantibody(concentrated).(TC-CodeAN). - DilutionBuffer –(whitecap)
1x20ml2.5xconcentrated(PBS,1%BSA,20mMEDTA,10KIU/mlaprotinin,20mMBenzamidine)(TC-CodeDC). - POXDilutionBuffer –(whitecap)
1x12mlPBS,1%BSAReadytouse(TC-CodeDD). - Substrate –(greencap)
1x12mlTMB(Tetramethylbenzidine)insubstratebuffercontainingH2O2.Readytouse(TC-CodeKN). - StopSolution –(redcap)
1x15ml0.5MSulphuricAcid(TC-CodeKK).Readytouse. - WashBuffer –(bluecap)
1x20ml12.5xconcentrated(PBS,0.5%,Tween20)(TC-CodeBE). - PlasminogenActivatorDetectionMixture
1xlyophilizedH-D-norleucyl-hexahydrotyrosyllysine-p-nitroanilidediacetatesaltandGluplasminogen(TCCodeBA). - DetectionMixtureDilutionBuffer (whitecap)
1x20ml50mMTris,12mMNaCl.Readytouse(TC-CodeDA).
MeasurementPrinciple:
IntheActibind®u-PAtestamonoclonalantibodywhichdoesnotblocku-PAfunctionalactivityiscoatedontoamicrotitreplateandusedtobindu-PAcontainedinplasmatotheplatesurface.Followinganincubationperiod,non-boundplasmacomponentsarewashedawayandadetectionmixturecontainingGlu-plasminogenandachromogenicplasminsubstrateisincubatedontheplate.Theboundu-PAactivatesGlu-plasminogentoyieldplasmin.Thereactionbetweenplasminandthechromogenicplasminsubstratereleasesacoloredproductwhoseconcentrationisproportionaltotheamountofactiveu-PAinthetestsample.Afterphotometricallymeasuringthisreaction,themicrotiterplateiswashedtoremovetheactivitysubstratesolution.Theu-PAantigenremainsboundtotheplate.AhorserADIshperoxidase(POX)conjugatedanti-u-PAantibodywhichrecognizesbothactiveu-PAandinactiveu-PAisthenincubatedontheplate.Followingincubationandwashingoftheplate,aPOXsubstrateisusedtoproduceacoloredreactionproductwhoseconcentrationisproportionaltothetotalu-PAcontentofthetestsample.
Background:
Humanurokinase(u-PA)isanenzymewhichfunctionsasanactivatorofthefibrinolyticenzymesystem.ItsABIlitytolysefibrinclotsmakesitusefulasaneffectivethrombolyticagentintheresearch ofavarietyofclinicaldiseasestatesincludingpulmonaryembolismandlocalizedthrombosis.Urokinaseissynthesizedbyandreleasedfromnumerouscelltypesincludingendothelialcells,kidneycellsandmacrophages.Severalmalignanttumors,especiallythoseoftheurogenitalandgastrointestinaltracts,havebeenshowntoproduceincreasedquantitiesofurokinase.
Urokinaseexistsinthreemajorforms:enzymaticallyinactivesinglechainurokinaseorpro-urokinase(scu-PA),enzymaticallyactivetwochainurokinase(tcu-PA)andurokinase-inhibitorcomplexes.scu-PAcirculatesinplasmaataconcentrationof1-2ng/mlandisconvertedtotcu-PAinvivobytheactionofplasminandkallikrein.Eachformofu-PAdisplaysadifferentactivity,differentaffinitiesforGlu-plasminogen,anddifferentratesofinhibitionbyplasmaproteaseinhibitors.
Forresearch admiNISTration,pro-urokinaseisgenerallypreferredoverotherformsofthisplasminogenactivator. Pro-urokinaseisnotinhibitedbyplasminogenactivatorinhibitorsand,althoughnoformofurokinaseishighlyfibrinspecific,pro-urokinaseismorefibrin-orientedthanotherformsofu-PA.Duringthrombolytictesting,pro-urokinaseisconvertedtotheactivetwochainurokinaseandisthensusceptIBLetoinhibition.TheActibind®u-PAassayemploysspecificreagentswhichallowforthemeasurementofurokinaseactivityoriginatingfromboththeactivetwochainformandthesinglechainformofthemolecule.Itdoesnotdetectinactiveurokinase-inhibitorcomplexes.u-PAantigen,however,isdetectedinallthreeforms(scu-PA,tcu-PAandu-PAinhibitorcomplexes)bythisassaysystem.Thisallowsthequantificationofurokinaseinhibitors.
Ifyouwishtodifferentiatebetweenpro-urokinaseandtwo-chainurokinase,theTechnozymscu-PAkitTC11110determinesonlysinglechainurokinase.
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通过解答,教会我,谢谢
请问下有无同学需要H37RA的?我是做EAE模型的,上个月购买了BDDifco公司的H37RA(货号),因为购买的时候只能整盒6支购买,但我们用不了那么多,所以想问问有无同学需要的,100mg/支,800元/支或用等价试剂交换。地址广州。有需要的请私信,谢谢!
(1)优级纯试剂 亦称保证试剂,为一级品,纯度高,杂质极少,主要用于精密分析和科学研究,常以GR表示。
(2)分析纯试剂 亦称分析试剂,为二级品,纯度略低于优级纯,杂质含量略高于优级纯,适用于重要分析和一般性研究工作,常以AR表示。
(3)化学纯试剂 为三级品,纯度较分析纯差,但高于实验试剂,适用于工厂、学校一般性的分析工作,常以CP表示。
(4)实验试剂 为四级品,纯度比化学纯差,但比工业品纯度高,主要用于一般化学实验,不能用于分析工作,常以 LR表示。
以上按试剂纯度的分类法已在我国通用。根据化学工业部颁布的“化学试剂包装及标志”的规定,化学试剂的不同等级分别用各种不同的颜色来标志,见表1。
表1 我国化学试剂的等级及标志
刚入这行,谢谢大家
我想检测血管组织中的钙离子浓度,不知道哪个公司有试剂盒
A.H2SO4Na2SO4AgNO3BaCl2.
B.NaOHNa2CO3NaHSO4MgCl2
C.CaCl2NaNO3MgSO4BaCl2
D.HNO3KOHKClK2CO3
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