Humanrhinovirus3Cprotease(HRV3CProtease)isacysteineproteasethatrecognizesthecleavagesiteofLeu-Glu-Val-Leu-Phe-Gln-Gly-Pro,commonlyreferredtoasthePreScissionSite.ItcleavesbetweenGlnandGly. TherecombinantformoftheHRV3Cproteaseisarestrictiongradeproteasethathasrobustactivityat4°Candhighspecificactivity.ThisHRV3CProteaseisa47kDaproteinwithbothGSTandHistagssoitcanberemovedbyeitherNi-chelatingorGlutathione(GSH)resin.AmutationinthemouseAmelxtri-tyrosyldomainresultsinimpairedsecretionofamelogeninandphenocopieshumanX-linkedamelogenesis.
ItisrecommendedtotesttheHRV3CProteasecleavagewithprotease-to-targetproteinratioof1:100(w/w)inthebuffersuitableforthetargetproteinat40°Covernight,withthetargetproteinconcentrationatorbelow1mg/ml.Underthiscondition,almost95%oftargetproteiniscleavedinmostcases.Theefficiencyofcleavagevariesduetothesequencesaroundthecleavagesite,theconformationandthesolubilityofthetargetprotein.IncludingreducingagentsuchasDTTorTCEPinthebufferhelpsthecleavage.Duetoitshighspecificity,moreproteasecanbeusedwithoutnon-specificcleavageoftargetproteins.Nonon-specificcleavageofcontrolcleavageproteinisobservedatprotease-to-controlcleavageproteinratioof1:10.
AdditionalInformation
| Synonyms | Turbo3C(HRV3C)Protease |
|---|---|
| Product# | H-1192 |
| Preparation | ItisrecommendedtotesttheHRV3CProteasecleavagewithprotease-to-targetproteinratioof1:100(w/w)inthebuffersuitableforthetargetproteinat4°Covernight,withthetargetproteinconcentrationatorbelow1mg/ml.Underthiscondition,>9 |
| StorageTemp | -20°C |
