I.U.B.:3.4.21.4

Trypsinisapancreaticserineproteasewithsubstratespecificitybaseduponpositivelychargedlysineandargininesidechains.Itisderivedfroma34kDainactiveprecursorzymogen,trypsinogen,afterenzymaticremovalofanN-terminal6-aminoacidleadersequenceresultinginthe23.8kDatrypsinmolecule.TheoptimumpHis8.0.Trypsinisinhibitedbyorganophosphoruscompoundssuchasdiisopropylfluorophosphateandnaturalinhibitorsfrompancreas.Soybean,limabean,andeggwhitearealsosourcesofnaturalinhibitors.TrypsincleavesamideandesterbondsofArgandLys.TheWorthingtonSequencingGradeTrypsinhasbeenfurtherpurifiedtoremovetracecontaminatingproteasesandautolysisproductswhichcouldinterfereintrypsindigestionexperiments,andexhibitsasinglebandonPAGE.

Uses:Fortissueculturework,Worthingtontrypsin,Codes:TRL,TRLS,TRLVMFandTRTVMFhavebeenusedbymanyresearchers.ProductCodes:TRSEQZ,TRSEQIIandTRTPCKaretypicallyusedforproteinsequencing,mappingandstructurestudies.Worthingtonmodifiedsequencinggradetrypsin,ProductCode:TRSEQZ,issubjectedtoextensivepurificationtoremovecontaminatingproteasesandtrypticautolysisby-productswhichcouldaffectthespecificityofthedigestionprocess. Subsequently,theenzymeischemicallymodifiedtominimizetheautolysisprocessaswellasincreasethestABIlity.Themodifiedtrypsinisprocessedfurthertoremoveresidualautodegradationproducts.Thespecificityoftheenzymeisroutinelycheckedafterthechemicalmodification.

Stability/Storage:MostgradesofWorthingtontrypsinarestablefor2-3yearswhenstoredat2-8°C.Protectfrommoisture.

UnitDefinitions:TAMEUnit:OneUnithydrolyzes1µmoleofp-toluene-sulfonyl-L-argininemethylester(TAME)perminuteat25°C,pH8.2,inthepresenceof10mMcalcium. 

Note:1mgtrypsin≥180TAMEunits,10,350BAEEunits,3,450USP/NFunits.

TechnicalNotes:TheVirusandMycoplasmaFreetrypsin(Code:TRTVMF)hasbeenfilteredthroughan0.22micronporesizemembrane,lyophilized,subjectedtogammairradiation,andtestedforvirusandmycoplasma.

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