细菌和酵母的粉剂培养基

蛋白酶抑制剂分类和应用在分离蛋白时，组织或细胞裂解液中充满了来自溶酶体的酶,并且原来无活性的蛋白酶原被充分激火,开始消化其周围的蛋白质.为避免此因素，蛋白酶抑制剂必须被加入在缓冲液内以阻止蛋白酶对细胞组织的消化作用.来自哺乳类动物，植物,霉菌或细菌的组织，蛋白酶成分是不同的,并根据机体状态的发展蛋白酶也可以发生变化.哺乳类动物组织分离时比细菌或其它组织分离时更需要应用蛋白酶抑制剂使组织细胞受到完全的保护.因此，在各种蛋白分离时， 在各种蛋白电泳时蛋白酶抑制剂的适当选用就成了很重要的因素。蛋白酶功能 :涉及到许多细胞内和细胞外的过程 :食物蛋白在消化道的消化激活无活性酶原和多肽激素/神经转移介质的释放o调节血凝物质和纤维素o内源性和外源性蛋白的细胞内消化跨膜的蛋白转移微生物对生物体攻击,通过释放蛋白酶的导致毒性增加蛋白酶分类 :根据催化中心结构的不同蛋白酶可分为 4类:丝氨酸蛋白酶在其活性中心含有丝氨酸和组氨酸，如:糜蛋白酶,弹性蛋白酶，血纤维蛋白溶酶，凝血酶，枯草杆菌蛋白酶，胰酶Chymotrypsin,Elastase,Plasmin,Subtilisin,Thrombin,Trypsin2.半胱氨酸蛋白酶（别名:硫醇或SH蛋白酶）在其催化中心含有半胱氨酸，如:Calpain,组织蛋白酶B,C和L，和木瓜蛋白酶3天(门)冬氨酸蛋白酶（羧基蛋白酶）在其活化中心含有天(门)冬氨酸，如:组织蛋白酶D,木瓜蛋白酶,肾素4.金属蛋白酶含有金属离子，即:锌2+在其催化中心，如氨基肽酶，羧基蛋白酶A和B，嗜热菌蛋白酶机体防止蛋白酶破坏作用几种可能性 :酶在特殊囊性小泡的分配(溶酶体，包涵体)内源性蛋白酶抑制剂（如:Aprotinin，抗凝血酶，抗胰酶）SERVA6 种蛋白酶抑制剂鸡尾酒的作用 :SERVA提供6种不同的蛋白酶抑制剂鸡尾酒，用于分离纯化各种组织的蛋白，多肽的最佳选择蛋白酶抑制剂混合物G含有5种水溶性的抑制剂用于抗丝氨酸-,半胱氨酸-和金属蛋白酶作用，并且是常用选择的鸡尾酒，应当避免同时使用有机体溶剂?

蛋白酶抑制剂混合物 HP含有4种水溶性抑制剂用于抗丝氨酸 -,半胱氨酸蛋白酶作用，并且有效地用于分离和提取重组蛋白， 避免金属螯合物。蛋白酶抑制剂混合物M含有6种不同的抑制剂用于抗丝氨酸-,半胱氨酸和天(门)冬氨酸酶以及金属蛋白酶作用，并用于哺乳动物组织的蛋白分离。蛋白酶抑制剂混合物P含有6种不同的抑制剂用于抗丝氨酸-,半胱氨酸和天(门)冬氨酸酶以及金属蛋白酶作用。此被推荐用于从植物来源的蛋白分离?

蛋白酶抑制剂混合物 FY含有4种不同的抑制剂用于抗丝氨酸 -,半胱氨酸,天(门)冬氨酸蛋和金属蛋白酶作用。用于从酵母和其它真菌来源的蛋白制备。蛋白酶抑制剂混合物B含有5种不同的抑制剂用于抗丝氨酸-,半胱氨酸,天(门)冬氨酸和金属蛋白酶作用。尤其应用于在细菌提取蛋白。产品包装:所有混合物为冻干粉剂.溶解在1ml水或DMSO浓度100x,足够用于100ml组织提取物每种抑制剂混合物的成分列表如下:成份混合G混合M混合P混合FY混合B混合HP391013910239103391043910539106AEBSF++++++Aprotinin++---+Bestatin-++-+-E-64++++++Leupeptin+++--+PepstatinA-++++-1,10-Phenanthroline--++--EDTA-Na2-salt+---+-注:AEBSF(PEFABLOCSC)和Aprotinin是丝氨酸蛋白酶的抑制剂苯丁抑制素(Bestatin)是氨基肽酶的抑制剂E-64是半胱氨酸蛋白酶的抑制剂Leupeptin是丝氨酸和半胱氨酸蛋白酶的抑制剂PepstatinA是天门冬氨酸蛋白酶的抑制剂EDTA和1,10-Phenanthroline是金属蛋白酶抑制剂蛋白酶抑制剂表 .以下表显示了竞争者产品与 SERVA蛋白酶抑制剂在应用或成分的比较SERVA

混合

G

混合M

混合P

混合FY

混合B

混合HP39101

39102

39103

39104

39105

39106S公司P2714P8340P9599P8215P8465P8849R公司

Complete

-

-

-

-

CompleteEDTA-freeC公司 539131 539134 - 539136 539132 539137蛋白酶抑制剂特征 :SERVA提供了全面的高纯度的用于4组蛋白酶(丝氨酸-,半胱氨酸和天(门)冬氨酸蛋白酶和金属蛋白酶)抑制剂的产品线以下应用表显示适用于不同领域抑制剂特征使用产品线正在伸展蛋白酶抑制剂的生产在制备中100多篇文献可查阅蛋白酶抑制剂应用范畴 :细胞调节过程的研究(如老年痴呆症/肿瘤研究)抗体,酶在诊断系统蛋白降解的抑制(如:诊断试剂盒)? 为了避免在细胞裂解时蛋白酶释放的破坏作用 (如:在制备性生化方面 )抑制由蛋白酶引起的组织裂解(如:细胞培养)应用表:此表总结了在生化和细胞生物学常见蛋白酶抑制剂的应用 (近40种).该表概括了作用机制,靶蛋白酶类型 ,溶解度,浓度和有关的文献参考货号 产品 分子量12745 AEBSF-HCl 239.7

作用/抑制的特异性溶解度浓度范围a)文献参考稳定性IrreversibleinhibitorofH2O,0.1-2Mμ60,83,84ThrombinandotherserineAqueousproteases.Inhibitsbysolutionsacylationoftheactivesiteofarestabletheenzyme.MuchlesstoxicbetweenpHthanPMSFandDFP5-612309Amastatin-HCl511Non-toxicreversiblemetallo-Ethanol1-100Mμ1,2,3,4proteaseinhibitor.Inhibits0.5%manymembrane-boundAcOHpeptidaseswhicharecriticalregulatorsofpeptidehormones,e.g.aminopeptidaseAandM,butnotaminopeptidaseB.Inhibitsalsoleucineaminopeptidase.1254813684

(epsilon)-131.2HighlyactiveinhibitorofH2O1-20Mμ5,6Aminocaproicfibrinolysinandacidchymotrypsin.(alpha)1-68000GlycoproteinthatinhibitsAqueousUsedat7,8,9,10Antichymotypsinchymotrypsin-likeproteasesbuffersequimolarfromhuman(aboveallhumanneutrophilconcentrationplasmacathepsinG)byformingstablecomplexes.Acutephaseprotein;concentrationinplasmaincreasesaftereventslikeinflammationortissuedamage13685Antipain-HCL678.2ReversibleinhibitorofserineH2O1-100Mμ11,12,13andcysteineproteases.MethanolInhibitspapainandtrypsinDMSOmorespecificlythanleupeptin.Plasminisinhibitedonlyslightly.AlsoinvolvedininhibitionofRNAsynthesis13695136941371813985139861452514980155201552115700

AntithrombinIIIca.Glycoproteinthatplaysafromhuman60000majorroleincontrollingplasmaserineproteasesinthebloodclottingcascade.Inactivatesaboveallthrombinbyforminganextremelystablecomplex,aneffectwhichisenhancedbyheparin.Inhibitsalsootherproteasesofthecoagulationcascadelikeplasmin,kallikrein,factorIXa,Xa,XIaandXIIa.(alpha)1-ca.GlycoproteinthatismainlyAntitrypsinfrom53000involvedinthecontrolofhumanplasmaneutrophilelastaseactivity.((alpha)1-Inhibitsalsomostofotherproteinasemammalianserineproteasesinhibitor)butatalowerrate.Blockstheactionoftargetenzymesbybindingnearlyirreversiblytotheiractivesite.Aprotinin(Trypsinca.Basicsingle-chaininhibitorfrom6500polypeptidethatinhibitsbovinelung)numerousserineproteasesbybindingtotheactivesiteoftheenzyme,formingtightcomplexes.Itinhibitsaboveallplasmin,kallikrein,trypsin,chymotrypsinandurokinase,butnotcarboxypeptidaseAandB,papain,pepsin,subtilisin,thrombinandfactorX.Usedincellculturetopreventproteolyticdamagetocellsandtoextendlifetimeofcells.ArphamenineA387.4Inhibitorofthemetallo-proteaseAminopeptidaseBArphamenineB403.4Inhibitorofthemetallo-proteaseAminopeptidaseBBenzamidine-HCl174.6PotentinhibitorofthrombinandtrypsinBestatin-HCl344.8Metalloproteaseinhibitorwithmulti-pharmacologicalfunctions.Inhibitscellsurfaceaminopeptidases(notablyB)andleucineaminopeptidase.InhibitorofleukotrieneA4hydrolaseandofenkephalindegradationincellpreparationsfrombrain.Hasanticarcinogenicandimmunomodulatingproperties.CA-074383.4InhibitorofCathepsinBCA-074-Me397.5ProinhibitorforintracellularCathepsinB.Membrane-permeableanalogofCA-074CalpainInhibitorI367.2Tripeptidealdehyde.SpecificinhibitoroftheCa2+-dependentcysteineproteasecalpainIandofcathepsinBandL.

H2O Usedat8,1415,16equimolarconcentrationsH2OUsedat8,9,17,18,19AqueousequimolarbuffersconcentrationsH2O,Incellculture:22,23,24,25Aqueous0.01-3g/ml;μbuffers.inotherSterileapplications:filtered10-250g/mlμsolutionsatpH5-8arestableforseveralmonths.DenaturesatpH>12H2O,MeOHIC50:85,860.006μg/mlH2O,MeOHIC50:85,860.002μg/mlH2O0.1-50Mμ26,27,28Methanol1-150Mμ29,30,31,32,33(5mg/ml)MitogeniceffectsatnmolarconcentrationsDMSO0.01-1Mμ87,88,89DMSO 1μM 90Ethanol,1-50Mμ34,35,36Methanol,DMF,DMSO(10mg/ml)1571052150171587720520600211002110221000210011128011290209472773651867

CalpainInhibitorII385.2Tripeptidealdehyde.SpecificinhibitoroftheCa2+-dependentcysteineproteasecalpainIIandofcathepsinBandL.Cathepsin489.5SpecificinhibitorofInhibitorZ-Phe-CathepsinB/L/SandPapainGly-NHO-Bz-pMeChymostatinca.600Peptide-derivedaldehyde(mixtureof3components).Reversibleinhibitorofchymotrypsin-likeserineandsomecysteineproteasesDFP184.2Apotentirreversibleinhibitor(Diisopropylfluoro-ofserineproteasesandphosphate)acetylcholineesterase.Highlytoxic!DiprotinA341.5Reversibleinhibitorofthemetallo-proteaseDipeptidylaminopeptidaseIVE-64357.4Non-competitiveirreversibleinhibitorofpapainandothercysteineproteases.Formsathioetherbondwiththesulfhydrylgroupintheactivecenteroftheenzyme.Usefulforactivesitetitration:onemoleofE-64inhibitsonemoleofprotease.E-64d(EST)342.4Membrane-permeableanalogofE-64cEbelactoneA338.5Non-toxicinhibitorforesterases,acylpeptidehydrolase,lipaseandN-formylmethionineaminopeptidaseEbelactoneB352.5Non-toxicinhibitorforesterase,lipaseandN-formyl-methionineaminopeptidase.InhibitsalsocarboxypeptidaseY-likeexopeptidase.EDTA-Na2372.3ReversibleinhibitorofmetalloproteasesEGTA380.4Inhibitsmetalloproteases..RevealshighselectivityforCa2+overMg2+ions.Elastatinal512.6InhibitorofElastaseLeuhistin241.3InhibitorofAminopeptidaseMLeupeptin-475.6Tripeptidealdehyde.hemisulfateReversiblecompetitiveinhibitorofserineandcysteineproteases.InhibitsalsophospholipaseDandCactivationinrathepatocytes.

Ethanol,1-50Mμ35,36,37,38Methanol,DMF,DMSO(10mg/ml)DMSO,Ki:91,92EtOH,0.15-16MμAcetonitrileDMSO10-100Mμ11,39,40,41AceticacidIsopropanol;10-100Mμ28,42,43aqueoussolutionsareunstableH2OKi:2.2Mμ94H2O,DMSO1-10Mμ51,52,53MixturesofwaterandethanolDMSO1μM90,99,100Methanol1-10Mμ43,44,45(200mg/ml)Methanol1-10Mμ44,46,47(200mg/ml)H2O(pH8)1-10Mμ48,49NH4OH,1-10Mμ50NaOHH2O,Ki:0.21Mμ11MeOH,(withAc-Ala-DMSOAla-Ala-OMeassubstrate)H2O,IC50:102,103,104DMSO,0.2μg/mlEtOH,MeOHH2O1-100Mμ11,56,57,58Stableforserevalmonthswhenstoredinaliquotsat-20 C°2830131682526823198532395327531701317016

(alpha)2-725000Glycoproteincomposedof4Macroglobulinidenticalsubunits.Broad-fromhumanrangeendoproteinaseplamasinhibitor.Inhibitsbyforminga?trap?aroundtheenzymeallowingonlysmallsubstratemoleculestoenterandtobecleavedbytheentrappedprotease.PEFABLOC?SC239.5Water-solubleandrelativlyb)</SUP<sup>non-toxicirreversible4-(2-Aminoethyl)-inhibitorofthrombinandotherserineproteases.benzenesulfonylInhibitsbyacylationofthefluorideactivesiteoftheenzyme.hydrochloridePepstatinA685.9Pentapeptidederivative.Reversibleinhibitorofasparticproteases,e.g.pepsin,cathepsinD,chymosin,reninPhebestin441.5InhibitorofAminopeptidaseNPMSF174.2IrreversiblyinhibitsserinePhenylmethylproteasesbysulfonylationofsulfonylfluoridetheserineresidueintheactivesiteoftheprotease.Inhibitsalsopapain(reversiblebyDTTtreatment)andacetylcholin-esterase.Doesnotinhibitmetallo-,aspartic-andmostcysteineproteases.Phosphoramidon587.5Specificinhibitorofthermolysinandneutralendopeptidase-24.11(ANPDegradationEnzyme).InhibitsalsotheactivityofEndothelinConvertingEnzyme,collagenaseandmetallo-endoproteinasesfrommanymicroorganisms.Doesnotinhibitserine,cysteineandasparticproteasesTLCK369.3Irreversiblyinhibitstrypsin(1-Chloro-3-butnotchymotrypsinbytosylamido-7-alkylatingthehistidineamino-2-residueintheactivesiteofheptanoneHCl)theenzyme.Inhibitsalsosomeotherserineandcysteineproteaseslikebromelain,ficinandpapain.TLCKdoesnotreactwithzymogensorinactiveprotease-inhibitorcomplexes.TPCK351.8Irreversiblyinhibits(1-Chloro-3-chymotrypsinbutnottrypsintosylamido-4-byspecificallyreactingwithphenyl-2-histidine.Inhibitsalsootherbutanone)serineandcysteineproteasesuchasbromelain,ficinandpapain.

H2OUsedat8,59Stableforequimolarseveralconcentrationsmonthswhenstoredinali