ProductOverview
Sonichedgehog(SHH)isusedtostudydifferentiationandexpansionofhumanandmouseembryonicandadultstemcells.StudiessuggestSHHisinvolvedinregulatingstemcellfatesofneuralandhematopoeticlineages1.Itcontrolscelldivisionofadultstemcellsandhasbeenimplicatedindevelopmentofsomecancers2.SHHisinstrumentalinearlyembryopatterninganddevelopment.Ithasbeenimplicatedasthekeyinductivesignalinpatterningoftheventralneuraltube,theanterior-posteriorlimbaxis,andtheventralsomites3.ThematureBIOLOGicallyactiveformofSHHmoleculeisproducedbyautocatalyticcleavageofitsprecursorproteinandcorrespondstoapproximatelytheN-terminalhalfoftheprecursormolecule.SonicHedgehogisanapproximately20kDaproteinexpressedinE.coliconsistingof175aminoacidresidues,includinganN-terminalIle-IlesequencesubstitutedforthenaturallyoccurringchemicallymodifiedCysresidue.
ProductSpecifications
Size
25µg
Source
E.coli
Formulation
Lyophilizedfroma0.22sterile-filteredsolutionin1xPBS &0.1MNaCl.
AminoAcidSequence
IIGPGRGFGKRRHPKKLTPLAYKQFIPNVAEKTLGASGRYEGKITRNSERFKELTPNYNPDIIFKDEENTGADRLMTQRCKDKLNALAISVMNQWPGVKLRVTEGWDEDGHHSEESLHYEGRAVDITTSDRDRSKYGMLARLAVEAGFDWVYYESKAHIHCSVKAENSVAAKSGG
UniprotAccessionNumber
Q15465,residues24-197
Purity
Greaterthan98%bySDS-PAGEanalysis
EndotoxinLevel
Lessthan0.1ng/µg(1EU/µg)ofSHHasdeterminedbytheLALmethod.
BiologicActivity
TheED50islessthan1μg/mlasdeterminedbyitsABIlitytoinducealkalinephosphataseproductionbyC3H10T1/2fibroblasts.
StorageandStability
StemfactorSHHisshippedatroomtemperature.LyophilizedSHHisstableforupto6monthsfromdateofreceiptwhenstoredat-20°Cto-80°C.ReconstitutedSHH,atconcentrationsgreaterthan0.1mg/ml,isstableforupto3monthswhenstoredat-20°Candfor1yearwhenstoredat-80°C.
Reconstitution
Centrifugebrieflyandthenreconstitutein1XPBStoyieldastocksolutionofnolessthan0.1mg/mlofSHH.Reconstitutedproteinshouldbeusedimmediatelyorstoredinworkingaliquotsat-20°C.Forlong-termstorage,itisrecommendedtoaddacarrierprotein(0.1%HSAorBSA).Avoidfreeze-thawcyclesasitcanresultinlossofactivity.
SpecificationSheets
- 03-0008ProductSpecificationSheet
SafetyDataSheets
- 03-0008SafetyDataSheet
References
- Briscoe,J.(2009)Makingagrade:SonicHedgehogsignalingandthecontrolofneuralcellfate.EMBOJ.28:457-465.
- Gough,N.M.,Gearing,D.P.,King,J.A.,Willson,T.A.,Hilton,D.J.,Nicola,N.A.,andMetcalf,D.(1988)Molecularcloningandexpressionofthehumanhomologueofthemurinegeneencodingmyeloidleukemia-inhibitoryfactor.Proc.Natl.Acad.Sci.USA85:2623-2627.
- Ingham,P.W.,andMcMahon,A.P.(2001)Hedgehogsignalinginanimaldevelopment:parADIgmsandprinciples.GenesDev15:3059-3087.
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