ORDERINGINFORMATION
CatalogNo.:13042
Format:100ugantigen-affinitypurifiedIgGinTris-citrate/phosphatebuffer,pH7-8,0.09%sodiumazide.
BACKGROUND
AurorakinaseA,alsoknownasserine/threonine-proteinkinase6,isanenzymethatinhumansisencodedbytheAURKAgene.AuroraAisamemberofafamilyofmitoticserine/threoninekinasesassociatedwithimportantprocessesduringmitosisandmeiosis.AuroraAisactivatedbyoneormorephosphorylations,anditsactivitypeaksduringtheG2phasetoMphasetransitioninthecellcycle.AuroraAdysregulationhasbeenassociatedwithhighoccurrenceofcancer.
SPECIFICATIONSUMMARY
Antigen:Syntheticpeptiderepresentingaportionoftheproteinencodedwithinexons5and6.
Accessionno.:NP_003591.2
GeneID:6790
HostSpecies:Rabbit
Specificity:ThisantibodyreactswithhumanAurorakinaseA,andtheepitoperecognizedmapstoaregionbetweenresidues75-125ofhumanAurorakinaseA.Basedon100%sequenceidentitythisantibodyispredictedtoreactwithorangutan,gorilla,chimpanzee,andNorthernwhite-cheekedgibbon.
APPLICATIONS
Immunoblotting:1:5,000-1:20,000dilution
DetectionofhumanAuroraAinHeLaandHEPG2wholecelllysateswithcatalogno.13042at0.2ug/ml.Detectionbychemiluminescence.
Immunoprecipitation:1-5ug/mglysate.
Thesearerecommendedconcentrations.Endusersshoulddetermineoptimalconcentrationsfortheirapplications.
DILUTIONINSTRUCTIONS
DiluteinPBSormediumthatisidenticaltothatusedintheassaysystem.
STORAGEANDSTABILITY
Thisantibodyisstableat2-8°Cforone(1)yearfromdateofreceipt.
Forinvitroinvestigationaluseonly.Notintendedforuseintherapeuticordiagnosticprocedures.
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2、可逆抑制剂:包括
a、竞争性抑制剂,抑制剂与底物竞争性结合酶反应中心,使Km增大,而Vmax不变 b、非竞争性抑制剂,酶与抑制剂结合后还能与底物结合,但活性降低,使Vmax减小,而Km不变
c、反竞争性抑制剂,酶只能与底物结合后才能与抑制剂结合,Vmax与Km都减小
可逆抑制剂可用透析等方法除去,使酶恢复作用
1、测定酶比活力:底物需要过量么?测定时间多长?是否可以加入过量的底物,然后测定3min吸光度的增加值,从吸光度的变化值计算比活力。
2、在酶抑制剂筛选的过程中,是否需要保证底物过量?还是要水浴一定时间让反应完全?我看到文献说用终浓度为0.1μmol/mL的底物,终浓度为45U/ml的酶,我想问,假设总体积为1ml,那么终浓度为45U/ml的酶岂不是每分钟能转化4.8μmol的底物?那么0.1μmol/mL的底物不就几秒钟就反应完了?那么怎么测定初速度?
《血管紧张素转换酶抑制剂在心血管病中应用的中国专家共识》.PDF(242.69k)
1.对于抑制剂筛选工作(求ic50)是不是体系内酶与底物的量(底物应该是过量的)对实验结果影响不大。
2.如果要求算Km值,是不是需要知道反应产物的绝对量。反应时间文献上都是5分钟,反应速度就用反应产物量除以反应时间即可。
3.酶是进口分装的,规格5U,一次用不完,用PBS稀释后如何保存
谢谢