Soluble in 50% acetic acid. Ki for HIV I and HIV 2 proteases is 20 nm and 5 nm, respectively, at ph 4.7.
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1 Inhibitor Target Protease/ Mechanism of Action Class Effective Concentrations Notes Stock Solutions Working Concentrations Acetyl Pepstatin M.W Aspartic/Reversible InhibitsHIV 1and HIV 2proteases nM Solublein50%aceticacid.Ki forhiv IandHIV 2proteases is20nmand5nm, respectively,atph4.7. 1mg/1.553mlof 50%aceticacid=1 mmsolution dilute1:1000to obtain1µm concentration AEBSF, hydrochloride M.W Serine/Irreversible Inhibitstrypsin, chymotrypsin, plasmin,thrombin, andkallikrein. <1mM SolutionsinH20arestablefor 1 2monthsat4 C,pH7.Will slowlyhydrolyzeatph>8. 50mg/2.09mlH2O =100mMsolution Useat1mMorless. Amastatin Streptomycessp M.W Metallo/Reversible Inhibitsalanylaminopeptidase. 1 10µM Aqueoussolutionsarestable for1day.stocksolutions1 mminetoharestablefor1 monthat 20 C. 1mg/2.1ml anhydrousetohor 5mg/10.5ml anhydrousetoh= 1mM Dilute1:100fora 10µMworking solution Antipain, hydrochloride M.W Serineand Cysteine/Reversible Inhibitspapainand trypsin.plasminis inhibitedtoasmall extent.morespecific thanleupeptinfor papainandtrypsin µM Stableforseveralhours.Stock solutionsof10mminh20or bufferarestablefor1weekat 4 C,1monthat 20 C.Also solubleinmeohanddmso 5mg/8.27mlH2O =10mM Dilute1:100fora 100µMworking solution AntithrombinIII HumanPlasma M.W.65,000 Serine/Reversible Inhibitsthrombin, factorxa,trypsin,and othertrypsin like serineproteases useatequimolar concentration Aqueousstocksolutionsare stableat 20 C. 1mgin17.24ml 150mMNaCl,100 mmsodiumcitrate, 10mMTris HCl,pH 8.3=1mM useat concentration equimolarwith protease
2 p APMSF Hydrochloride MW252.7 Serine/Irreversible Inhibitstrypsin like serineproteases.no effectonacetylcholine esterase µM Mustbepreparedfresh.Halflifeof6mininpH buffersystems.stock solutionsof50mminh2oare stablewhenaliquotedand storedat 20 C. 5mg/346µlinH2O =50mM Dilute1:500fora 100µMsolution. Aprotinins BovineLung M.W.6512 Serine/Reversible Inhibitsserine proteases,including plasmin,kallikrein, trypsin,and chymotrypsin.does notinhibitthrombinor factorxa µg/ml Verystable.InactiveatpH> 12.8.Verysolubleinwater10 mg/ml.aliquotsarestableat 20 C. 10mg/mlinPBS Dilute1:5000fora 2µg/mlfinal concentration ArphamenineA Chromoboderium violoceum M.W Metallo Ahighlyspecific inhibitorof aminopeptidaseb nM Solubleinwater.Stock solutionsarestableforupto1 monthat 20 C 1mg/2.58mlH20= 1mMsolution Dilute1:1000to obtain1µm concentration. Bestatin M.W Metallo/Reversible Inhibitsalanylaminopeptidase 1 10µM Stablefor1day.Stock solutionsof1mmmeohare stableforatleast1monthat 20 C. 2mg/6.48ml MeOH=1mM Dilute1:100for10 µmworking solution. Calpeptin M.W Cysteine Cell permeable inhibitorofcalpaini andiiandpapain µM Acell permeablecalpain inhibitor.solubleindmso anddmf.diluteinaqueous mediumjustbeforeuse. 5mg/13.8ml DMSO=1mM solution Dilute1:1000to obtain1µm concentration. Cathepsin InhibitorI M.W Cysteine/Irreversible InhibitscathepsinB, cathepsinl,cathepsin S,andpapain µM Protectfromlight.Half lifein abufferedsolution(ph5 9) is25hoursat30 Candabout 2weeksat 20 C. 1mg/210µlDMSO oretoh=10mm solution Dilute1:50to obtain200µm concentration Cathepsin InhibitorII M.W Cysteine/Irreversible InhibitscathepsinB, cathepsinl,cathepsin Sandpapain µM Protectfromlight.Half lifein abufferedsolution(ph5 9) is27hoursat30 Candabout 2weeksat0 C 1mg/204µlDMSO oretoh=10mm solution concentration. Dilute1:50to obtain200µm
3 Cathepsin InhibitorIII M.W Cysteine/Irreversible InhibitscathepsinB, cathepsinl,cathepsin S,andpapain µM ProtectfromlightHalf lifein abufferedsolution(ph5 9) is37hoursat30 Candabout 2weeksat0 C. 1mg/198µlDMSO oretoh=10mm solution concentration. Dilute1:50to obtain200µm Cathepsin/Subtil isininhibitor M.W CysteineandSerine/ Irreversible InhibitscathepsinL, subtilisin,and thermitase µM Protectfromlight.Half lifein abufferedsolution(ph5 9) is2.5hoursat30 Candabout 40hoursatO C. 1mg/193µlin DMSOorEtOH=10 mmsolution Dilute1:100to obtain100µm concentration Chymostatin CysteineandSerine/ Reversible Inhibitschymotrypsinlikeserineproteases including,andα,β,γ, andδ chymotrypsin Stableforseveralhours. SolubleinDMSOandglacial aceticacid.solutionsare stableformonthsat 20 C 5mg/825µlin DMSO=10mM solution Dilute1:100fora 100µMworking solution. Cystatin,Egg White M.W.12,700 Cysteine/Reversible Inhibitscysteine proteases,including dipeptidylpeptidasei andiii,papain.ficin. andcathepsinb. Useatequimolar concentrations. Verystable.Stabletoheat. Freezeinthepresenceof20% glycerolorbufferedtoph7.5 Atleast1mg/mlin 20mMTris,pH7.5 Useatequimolar concentration. 3,4 Dichloroisocoumarin M.W Serine/Irreversible Inhibitsawiderange ofserineproteases. Notactivetowards beta lactamases µM Half lifeof20minutesatph 7.5.Stocksolutionsof10mM indmfordmsoarestablefor monthsat 20 C. 10mg/4.65mlin DMSO=10mM solution Dilute1:100for 100µMworking solution DFP M.W Serine/Irreversible Inhibitsserine proteases.inactivates acetylcholinesterase 100µM Verytoxic:specialhandling required.half lifeof1hourat ph7.5.stocksolutions Mindrypropanolstable forseveralmonthsat 70 C. 1g/10.86mlindry propanol=0.5m Dilute1:5000for 100µMworking solution
4 Dipeptidylpeptid aseivinhibitori M.W Serine/ProlineSpecific Inhibits dipeptidylpeptidaseiv µM DMSOstocksolutionsare stableforupto6monthsat 20 C.Useonlyhighquality, moisturefree,dmso.aqueous solutionsarestablefor1day. 1mg/219.5µl DMSO=10mM solution Dilute1:100to obtaina100µm workingsolution. Dipeptidylpeptid aseivinhibitor II M.W Serine/Proline Specific/Irreversible Acylatinginhibitorof dipetidylpeptidaseii andiv 1 10µM Protectfromlight.Half lifein abufferedsolution(ph7.6)is 24hoursat30 C. 1mg/2.81ml DMSOorEtOH=1 mmsolution. Dilute1:100to obtain10µm concentration DiprotinA M.W Metallo/Reversible 10 50µM Stablefor1dayatworking concentrations. Inhibitsdipeptidylpeptidase IV.Stocksolutionscanbe prepared1mminwater, MeOH.orEtOH.Stabilityhas notbeendetermined. 2mg/5.86ml=1 mmsolution Dilute1:20fora50 µmworking solution EST M.W Cysteine/Irreversible Cell permeable inhibiloroflysosomal cysteineproteases µg/ml Amembrane permeable calpaininhibitor.stock solutionsarestableforupto6 monthsat 20 C. 1mg/mlinEtOH Use20to40µlper mlofmedium E 64 Protease Inhibitor M.W Cysteine/Irreversible Inhibitscysteine proteases.doesnot affectcysteineresidues inotherenzymesor reactwithlow molecularweight thiolssuchasbetamercaptoethanol,very specific,activesite titrant. 1 10µM StablefordaysatneutralpH. Stocksolutionsof1mMin aqueoussolutionsarestable formonthsat 20 C. 5mg/13.99ml=1 mmsolution Dilute1:100fora 10µMworking solution.
5 EbelactoneB Streptomycessp. M.W Serine Aspecificinhibitorof carboxypeptidase Ylikeexopeptidase 0.1 2mg/ml SolubleinMeOH(200 mg/ml),etohand chloroform.solutionsare stablefor1weekat3 Cand1 monthat 20 C. 1mg/2.84ml=1 mmsolution Use2.84or5.08 µl/mlfor1or2ug, respectively. Ecotin,E.coli M.W.32,200 Serine Abroadrangeserine proteaseinhibitor. Inhibitschymotrypsin, elastase,factorsxa andxiia,andtrypsin 0.5to1.5nM Avoidfreeze/thawcycles. Storethestocksolutionat 70 C. Providedasa solutionin1mm HCl checkforlot specificvolume informationand diluteaccordingly EDTA,Disodium salt,m.w Mayinterferewith othermetal dependent biologicalprocesses 1 10mM stableinaqueoussolution. Stocksolutionsof0.5Min H2OatpH8.5arestablefor severalmonthsatrt. 1.9g/10ml=0.5M solution Dilute1:50fora10 mmworking solution EDTA, Tetrasodium salt,m.w Metallo/Reversible chelator Mayinterferewith othermetal dependent biologicalprocesses 1 10mM stableinaqueoussolution. Stocksolutionsof0.5MinH2O atph8.5arestablefor severalmonthsatrt. 1.9g/10ml=0.5M solution Dilute1:50fora10 mmworking solution Elastatinal M.W Serine/Reversible Inhibitselastaseand elastase likeserine proteases µM Stableforseveralhours.Stock solutionsof10mminh20are stablefor1weekat4 Cand forseveralmonthsat 20 C. 5mg/975µl=10 µmsolution Dilute1:100fora 100µMworking solution. leuhistin M.W Metallo/Reversible Competitiveinhibitor ofaminopeptidasem 1 2µg/ml Solubleinwater,EtOH,and DMSO.Storestocksolutions at 20 C. 1mg/mlofH2O Use1to2µl/ml.
6 leupeptin, Hemisulfate MW475.6 Serineand Cysteine/Reversible Inhibitstrypsin like proteasesandsome cysteineprotease; including endoproteinaselys C, kallikrein,papain, thrombin,cathepsinb, andtrypsin µM Stableforseveralhours.Stock solutionsof10mmonh2o arestablefor1weekat4 C and1monthat 20 C. 5mg/1.054ml=10 mmsolution Dilute1:100fora 100µMworking solution α2 Macroglobulin, HumanPlasma M.W.725,000 BroadRange/ Irreversible Inhibitsbromelain, chymotrypsin, elastase, endoproteinase,glu C, ficin,papain,plasmin, subtilisin,thermolysin, thrombin,andtrypsin. Forms"trap"around mostproteases. Useatequimolar concentrations. Aqueousstocksolutionsare stablefor6monthsat 20 Cat ph6 7.5.Sensitivetoacidic ph;denaturedbelowph4.0. Ammonia,methylamine,and hydroxylamine(aboveph7. 0)causeirreversible conversiontotheclosedform. Donotuseinthepresenceof DTT. 1mg/13.8µl=100 µmsolution Useatequimolar concentrations. NCO 700 M.W Cysteine Inhibitscalpain, cathepsinb,cathepsin L,andpapain mM Prepareasolutionontheday ofuse.inaqueoussolution, thismaterialisstableforup to6hoursatroom temperature.stableforlonger periodsinetoh. 1mg/87.6mlPBS. H,OorEtOH=10 mmsolution Dilute1:100to obtaina100µm workingsolution. Pepstatin M.W Aspartic/Reversible InhibitscathepsinD, cathepsing,pepsin, renin,andmany microbialaspartic proteases.weak bindingtoproteases. 1µM SolubleinMeOHto1mg/ml. Solubleto1m/mlinEtOhHif allowedtositovernightand to333mg/mlin6nacetic acid.stablefor1weekat4 C 5mg/7.29ml=1 mm Dilute1:1000fora 1µMworking solution
7 Phosphoramidon,disodiumsalt M.W Metallo/Reversible Inhibitorofsomemeta lloendopeptdases.a highlyspecific inhibitorof thermolysin 25mg/ml(8.5 mm) Stocksolutionsof1mg/mlin H2Oarestableforatleast1 monthat 20 C. 1mg/1.015ml= 1.7mMsolution Dilute1:200fora 8.5µMworking solution. PMSF M.W Serine/lrreversible Inhibits carboxypeptidasey, chymotrypsin,factor Xa,papain,plasmin, proteinasek, subtilisin,thrombin, andtrypsin mM Toxic:Mustbepreparedfresh andaddedatseveralsteps duringsamplepreparation. Half lifeof1houratph7.5. Stocksolutionsof200mMin anhydroussolvents(meoh, EtOH)arestableforatleast9 monthsat4 C. 100mg/2.87ml= 200mMsolution Dilute1:200fora1 µmworking concentration. Subtilisin InhibitorI M.W Serine Inhibitssubtilisinand thermitase µM Protectfromlight.Half lifein abufferedsolution(ph5 9) is4hoursat30 C,60hoursat 0 C. 1mg/264µlDMSO oretoh=10mm solution. Dilute1:100to obtain100µm concentration. Subtilisin InhibitorII M.W Serine Inhibitssubtilisinand thermitase µM Protectfromlight.Half lifein abufferedsolution(ph5 9) is4hoursat30 C,60hoursat 0 C. 1mg/210.3µl DMSOorEtOH=10 mmsolution. Dilute1:100to obtain100µm concentration. Subtilisin InhibitorIII M.W Serine Inhibitssubtilisinand thermitase µM Protectfromlight.Half lifein abufferedsolution(ph5 9) is8hoursat30 C,120hours at0 C. 1mg/198µlDMSO oretoh=10mm solution. Dilute1:100to obtain100µm concentration. Subtilisin InhibitorV M.W Serine/Cysteine/Irrev ersible Inhibitssubtilisinand elastase µM Protectfromlight.Half lifein abufferedsolution(ph5 9) is4hoursat30 C,60hoursat 0 C. 1mg/181µlDMSO oretoh=10mm solution. Dilute1:100to obtain100µm concentration.
8 TLCK Hydrochloride M.W Serine/Irreversible Inhibitstrypsin like serineproteases includingbromelain, endoproteinasearg C, endoproteinaselys C, ficin,papain,plasmin, thrombin,andtrypsin µM VeryunstableabovepH7.5. Stocksolutionsof10mMin aqueoussolutions(1mmhci, ph3.0)ormeohshouldbe preparedfreshasneeded. 5mg/1.354ml=10 mmsolution Dilute1:100fora 100µMworking solution TPCK M.W Serine/lrreversible Inhibitschymotrypsinlikeserineproteases includingbromelain, chymotrypsin,ficin andpapain µM Stableforseveralhours.Stock solutionsof10mminmeoh arestableforseveralmonths at4 C. 5mg/1.42ml=10 mmsolution Dilute1:100fora 100µMworking solution TrypsinInhibitor Soybean M.W.20,000 Serine/Reversible InhibitsfactorXaand trypsin Useatequimolar concentrations. DissociatedatlowpH.Stock solutionsindilutebuffersare stableat 20 C. 20mg/ml=1mM solution Useatequimolar concentrations
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