Type
Recombinant
Description
MembersoftheHedgehog(Hh)familyarehighlyconservedproteinsthatarewidelyrepresentedthroughouttheanimalkingdom.ThethreeknownmammalianHhproteins,Sonic(Shh),Desert(Dhh)andIndian(Ihh),arestructurallyrelated,andshareahighdegreeofaminoacidsequenceidentity(e.g.ShhandIhhare93%identical).TheBIOLOGicallyactiveformofeachHhmoleculeisobtainedbyautocatalyticcleavageoftheirprecursorproteins,andeachcorrespondstoapproximatelyonehalfoftheN-terminalportionoftheprecursormolecule.AlthoughHhproteinshaveuniqueexpressionpatternsanddistinctbiologicalroleswithintheirrespectiveregionsofsecretion,theyusethesamesignalingpathwayandcanbesubstitutedforoneanotherinexperimentalsystems.RecombinantMurineSonicHedgehogisa20.0 kDaproteinconsistingof176 aminoacidresidues,includinganN-terminalIle-Val-Ilesequencesubstitutedforthenaturallyoccurring,chemicallymodifiedCysresidue.
AminoAcidSequence
IVIGPGRGFGKRRHPKKLTPLAYKQFIPNVAEKTLGASGRYEGKITRNSERFKELTPNYNPDIIFKDEENTGADRLMTQRCKDKLNALAISVMNQWPGVKLRVTEGWDEDGHHSEESLHYEGRAVDITTSDRDRSKYGMLARLAVEAGFDWVYYESKAHIHCSVKAENSVAAKSGG
Source
E.coli
Purity
90%
BiologicalActivity
DeterminedbyitsABIlitytoinducealkalinephosphataseproductionbyC3H/10T1/2(CCL-226)cells.TheexpectedED50forthiseffectis0.5–1.0 μg/ml.
Endotoxin
Endotoxinlevelis<0.1 ng/μgofprotein(<1EU/μg).
Reconstitution
Centrifugethevialpriortoopening.Reconstituteinwatertoaconcentrationof0.1–0.5 mg/ml.Donotvortex.Forextendedstorage,itisrecommendedtofurtherdiluteinabuffercontainingacarrierprotein(example0.1%BSA)andstoreinworkingaliquotsat-20°Cto –80°C.
Storage/Expiration
–20°C
