Haematologic Technologies/FPR-chloromethylketone (PPACK)/FPRCK-01/5 mg, 25 mg, 100 mg

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货号:FPRCK-01
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品牌:haemtech
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商品描述
FormulationLyophilized
MolecularWeight524
Storage4°C
PurityHPLCandTLCanalysis
CompoundFPR-CK.2HCl
Assay
ShelfLife(properlystored)12months
ChemicalFormulaC21H33O3N6Cl3
StructureofBiotinylatedChloromethylketones
ThestructureofbiotinylatedFPRchloromethylketones(BFPRCK)(TOP)andbiotinylatedEGRchloromethylketone(BEGRCK)(BOTTOM)areshown.TheSPACERrepresentacarbon-spacerusedtooptimizethereactivityofthebiotin-groupaftertheprobehasbeenreactedwiththeactivesiteofaserineprotease.

Overview:

Tri-peptidechloromethylketoneshavebeenutilizedextensivelytoirreversIBLyinhibitvariousserineproteases(1-5).AmongthemostcommonchloromethylketonesareFPRCK(Phe-Pro-Arg-chloromethylketone;commonlyreferredtoasPPACK),whichisarapidthrombininhibitorandEGRCK(Glu-Gly-Arg-chloromethylketone;commonlyreferredtoasGGACK),whichisarapidfactorXainhibitor(1).BothFPRCKandEGRCKareusedextensivelyduringproteinisolationprocedurestoinhibitserineproteaseactivityandpreventfurtherconversionofzymogenstoactiveenzymes.Recently,themodificationofthesetri-peptidechloromethylketoneswithreportinggroups,suchasfluorescentprobes(6-8,14),rADIoactivelabels(9)orthioreactive-labels(10),hasprovidedauniqueapproachtothestudyofvariousserineproteases.Theseprobesareusefulbecausetheyallowameansofreportingmolecularchangesinanenzyme,andnotitszymogen,whilealsoinhibitingtheenzymaticactivity.

TheuseofbiotinasareportinggrouphasbeenusedextensivelywithantibodiesinELISAbasedassaysandinwesternblotting.Thebiotin,inconjunctionwithavidin,createsahighlysensitivemethodfordetectingantibodies,andtherefore,antigens.Bymodifyingthetripeptide-chloromethylketoneswithabiotingroup,thesensitivityoftheavidin/biotinsystemcanbeextendedtostudyserineproteaseswithouttheneedforspecificantibodiestotheactiveenzymes.

Biotinylatedtripeptidechloromethylketonescanbeusedinavarietyofways(11-13).First,thecompoundscanbereactedwithunwantedserineproteasesinasampleorpreparation,andcanthenberemovedalongwiththeproteaseusingavidin-Sepharose(11).Second,thebiotinylated-serineproteasecanbevisualizedonablotwithouttheuseofspecificantibodies(11).Third,thebiotinylatedserineproteasecanbequantitatedinanactive-sitespecificimmunoassay(12,13,15),suchasthetPA-CASSIA(seeAssayKits).ThespacerutilizedonthesecompoundshasbeenoptimizedtoallowgoodreactivityofthebiotinylatedFPRCKandthebiotinylatedEGRCKintheabovementionedprocedures.

Inadditiontobiotinylatedchloromethylketones,fluoresceinlabelledcompoundsarealsoavailable.ThefluoresceinlabelledcompoundsareusefulinbothWesternblotandfluorescentimagingapplications.

BiotinylatedandfluoresceinlabelledFPRCKandEGRCKarepreparedbythemethodofWilliamsetal.(11).

Properties:

SpecialPropertiesTri-peptidechloromethylketones(CMK)areverypotentandirreversibleinhibitorsofserineproteases.BFPRCKisespeciallyusefulforinhibitionofthrombinandtPA,whileBEGRCKisusefulforinhibitionoffactorXa.ThebiotinmoietyprovidestheABIlitytousethepeptide-CMKsasspecificprobesfordetectionand/orcaptureofserineproteasesviatheavidin/biotininteraction.
SpecialNotesFPRCKandEGRCKaresuppliedlyophilized,andshouldbestoredat4oC.BiotinylatedCMKsaresuppliedin10mMHClandshouldbestoredfrozedat-20oCorcolder.FluorosceinCMKsaresuppliedinDMSO,andshouldalsobestoredat-20oCorcolder.
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