ORDERINGINFORMATION
Catalogno.:13060(cloneGTS-1)
Format:100uglyophilizedpurifiedantibodyin10mMPBS,pH7.4,1%BSA.
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:MicrosomalfractionoftransformedmouseT-celltransfectedwithrathemeoxygenase-1(WR19LrHO-1).
Accessionno.:NP_036712
GeneID:24451
HostSpecies:Mouse
AntibodyClass:IgG1
Specificity:Thisantibodyrecognizeshuman,rat,andmouseHO-1.ItdoesnotrecognizerabbitHO-1anddoesnotcross-reactwithHO-2.
APPLICATIONS
Immunoblotting:useat5-10ug/ml.Abandof~33kDaisdetected.
DetectionofHO-1inhumansmallintestine.
DetectionofratHO-1with#13060at5ug/ml.
Immunohistochemistry:useat5-10ug/mlonhumanfrozenorparaffin-embeddedtissue.Antigenretrievalwith0.4ug/mlProteinaseK.
Thesearerecommendedconcentrations.Endusersshoulddetermineoptimalconcentrationsfortheirapplications.
DetectionofHO-1inhumanpancreas.
DetectionofHO-1inhumanpancreaticcarcinoma.
DetectionofHO-1inhumanthyroidcarcinoma.
DetectionofHO-1inhumanlungcarcinoma.
RECONSTITUTIONINSTRUCTIONS
Dissolvelyophilizedantibodyin50ulofdistilledwater;finalconcentrationwillbe2mg/ml.Iffurtherdilutionisneeded,dilutethisstockwith10mMPBS,pH7.4+1%BSAimmediatelybeforeuse.
STORAGEANDSTABILITY
Stocksolutionof2mg/mlshouldbestoredforone(1)yearat-20oCinappropriatealiquotstoavoidmultiplefreeze-thawcyclesorat4oCfor6monthswiththeadditionof0.1%sodiumazide.Dilutedantibodyshouldnotbestored.
Forinvitroinvestigationaluseonly.Notintendedfortherapeuticordiagnosticprocedures.
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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.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
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)
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