ORDERINGINFORMATION
Catalogno.:13061(clone1F12.A6)Format:100uginPBS(1mg/ml),pH7.4.PurifiedbyProteinGaffinitychromatography.BACKGROUND
Heme-oxygenaseisanenzymethatcatalyzeshemecatabolismtoyieldbiliverdin,iron,andcarbonmonoxide(CO).Therearethreeisoformsofheme-oxygenase:HO-1,HO-2,andHO-3.HO-1andHO-2havebeenidentifiedasthetwomajorisoformsinmammals.HO-1,alsoknownasheatshockprotein32(Hsp32)isinducedbymostoxidativestressinducers,cytokines,inflammatoryagents,andheatshock.HO-1deficiencyappearstocausereducedstressdefense,apro-inflammatorytendency,susceptibilitytoatheroscleroticlesionformation,endothelialcellinjury,andgrowthretardation.Therefore,up-regulationofHO-1isoneofthemajordefensemechanismsagainstoxidativestress.
SPECIFICATIONSUMMARY
Antigen:Syntheticpeptidecorrespondingtoaa1-30ofhumanHO-1.Accessionno.:P09601,NP_002124.1HostSpecies:MouseAntibodyClass:IgG1Specificity:Thisantibodyrecognizeshuman,mouse,andbovineHO-1.Itdoesnotcross-reactwithHO-2.APPLICATIONS
ELISA:useat1ug/mlwithHO-1onthesolidphase.
Immunoblotting:useat0.5-1ug/ml.Abandof~32kDaisdetected.
Immunofluorescence:useat5-10ug/ml
DetectionofHO-1informaldehyde-fixedHeLacells.
Positivecontrol:HumancelllinelysatessuchasA431,A549,HeLa,HepG2,Jurkat,orMCF7.
Thesearerecommendedconcentrations.Endusershoulddetermineoptimalconcentrationsfortheirapplications.
DILUTIONINSTRUCTIONS
DiluteinPBSormediumthatisidenticaltothatusedintheassaysystem.
STORAGEANDSTABILITY
Thisantibodyisstableforatleastone(1)yearat-20oC.Avoidrepeatedfreeze-thawcycles.
Forinvitroinvestigationaluseonly.Notintendedfordiagnosticortherapeuticapplications.
ebiomall.com
>
>
>
>
>
>
>
>
>
>
>
>
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)
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.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