Background
MembersoftheHedgehog(Hh)familyarehighlyconservedproteinswhicharewidelyrepresentedthroughouttheanimalkingdom.ThethreeknownmammalianHhproteins,Sonic(Shh),Desert(Dhh)andIndian(Ihh)arestructurallyrelatedandshareahighdegreeofamino-acidsequenceidentity(e.g.,ShhandIhhare93%identical).TheBIOLOGicallyactiveformofHhmoleculesisobtainedbyautocatalyticcleavageoftheirprecursorproteinsandcorrespondstoapproximatelytheN-terminalonehalfoftheprecursormolecule.AlthoughHhproteinshaveuniqueexpressionpatternsanddistinctbiologicalroleswithintheirrespectiveregionsofsecretion,theyusethesamesignalingpathwayandcansubstituteforeachotherinexperimentalsystems.RecombinantE.coliderivedHumanSonicHedgeHogisa20.0kDaproteinconsistingof176aminoacidresidues,includinganN-terminalIle-Val-IlesequencesubstitutedforthenaturaloccurringchemicallymodifiedCysresidue.
Specifications
Preparation
BiologicalactivitywasdeterminedbyitsABIlitytoinducealkalinephosphataseproductionbyC3H/10T1/2(CCL-226)cells.TheexpectedED50 forthiseffectis0.8-1.0μg/ml.Location
IVIGPGRGFGKRRHPKKLTPLAYKQFIPNVAEKTLGASGRYEGKISRNSERFKELTPNYNPDIIFKDEENTGADRLMTQRCKDKLNALAISVMNQWPGVKLRVTEGWDEDGHHSEESLHYEGRALDITTSDRDRSKYGMLARLAVEAGFDWVYYESKAHIHCSVKAENSVAAKSGGSource
E.coliReactivity
n/aTestedApplication
n/aStorage
ThelyophilizedSHHrecombinantproteinisstableforatleast2yearsfromdateofreceiptat-20ËšC.ReconstitutedSHHisstableforatleast3monthswhenstoredinworkingaliquotswithacarrierproteinat-20ËšC.Aswithanyprotein,exposingSHHrecombinantproteintorepeatedfreeze/thawcyclesisnotrecommended.Whenworkingwithproteinscareshouldbetakentokeeprecombinantproteinatacoolandstabletemperature.SpeciesReactivity
n/aPurity
Greaterthan98%bySDS-PAGEgelandHPLCanalyses.
Endotoxinlevelislessthan0.1ngperμg(1EU/μg).