Cataloguenumber
ENZ-1032
ShippingConditions
HeparanaseActiveEnzymeisshippedfrozenondryiceunlessstatedotherwisebythecustomer.
ShippingfeestoN.AmericaandW.Europeis$250
ShippingfeestoAsia,AustraliaandE.Europeis$350
Introduction
Heparanaseisanendoβ-D-glucuronidase,whichdegradesheparansulfatesidechainsofheparansulfateproteoglycans(HSPGs)intheextracellularmatrix.HeparanaseplaysanimportantroleinECMdegradation,facilitatingthemigrationandextravasationoftumorcellsandinflammatoryleukocytes(1,2,3).Upondegradation,heparanasereleasesgrowthfactorsandcytokinesthatstimulatecellproliferationandchemotaxis(4,5).Heparanaseisaheterodimercomprisedofa50kDasubunitharboringtheactivesiteanda8kDasubunit.Itisproducedasalatent65kDaprecursorandproteolyticallyprocessedtoitsactiveform(1,6).Heparanaseishighlyexpressedinmyeloidleukocytes(i.e.neutrophils)inplateletsandinhumanplacenta.Humanheparanasewasfoundtobeupregulatedinvarioustypesofprimarytumors,correlatinginsomecaseswithincreasedtumorinvasivenessandvascularityandwithpoorProspectivesurvival(7,8).
Description
HeparanaseActiveEnzyme isproducedinCHOcells.
Theproteinispurifiedbyseveralorthogonalchromatographysteps.
Formulation
HeparanaseActiveEnzymeissuppliedin20mMAcetatebufferand750mMNaClpH5.4.
StorageProcedures
Storeat–80ºC,avoidrepeatedfreeze-thawcycles.
Purity
Greaterthan95.0%asdeterminedby:
(a)AnalysisbyRP-HPLC.
(b)AnalysisbySDS-PAGE.
Specificity
HeparanaseActiveEnzyme isidentifiedbyWesternblotanalysiswithpolyclonal
rabbitanti-HPA1antibodiesas2subunitsof8-kDaand50-kDa.
BIOLOGicalActivity
ThespecificactivityofHeparanaseActiveEnzyme in-housestandardisabout0.7 Units(1unit=1μmoleofreducingendsofheparansulfate substrateformedperminutepermgHeparanaseActiveEnzyme at37°C).The enzymaticactivityofeachHeparanaseActiveEnzyme batchiscomparabletothe standardasdeterminedbyactivityassayinwhichimmobilized heparan,releasedduetoheparanaseactivity,isquantified colorimetrically.Recommendedreactionbuffer:20mMCitrate Phosphatebuffer,pH5.4;50mMNaCl;1mMCaCl2.
References
1.Vlodavsky,I.etal..,Nat.Med.5(1999)793–802.
2.Vlodavsky,IandFriedman,Y.,J.Clin.Invest.108(2001)341–347.
3.Parish,C.R.etal.,Biochem.Biophys.Acta1471(2001)M99–M108.
4.Vlodasvsky,I.etal.,CancerMetastasisRev.9,(1990)203-226.
5.Bashkin,P.etal.,.Biochemistry28(1989),1737-1743.
6.Fairbanks,M.B.etal.,J.Biol.Chem.274(1999)29587–29590.
7.Koliopanos,A.etal.,CancerRes.61(1992)4655–4659.
8.Gohji,K.etal.,Int.J.Cancer95(2001)295–301.
DataSheet
ToviewtheFULLVERSIONdatasheetclick Here
Usage
ProSpec"sproductsarefurnishedforLABORATORYRESEARCHUSEONLY.Theproductmaynotbeusedasdrugs,agriculturalorpesticidalproducts,foodadditivesorhouseholdchemicals.
