Manufacturedfor:
Collagenase
Cat.No.:17100TypeI
17101TypeII
17102TypeIII
17103HepatocyteQualified
17104TypeIV
Sizes:Seecatalog.
Custompacksizesavailableuponrequest.
Storage:2to8°C(-5to-20°Cafterreconstitution)
Avoidmoistureandexposuretolight.
Avoidinhalationandskincontact.
Thisproductisintendedforcellortissuedisaggregationonly.
Background
Collagenase(fromclostridiumhistolyticum)isaproteasewithaspecificityforthebondbetweena
neutralaminoacid(x)andglycineinthesequencePro-X-Glyc-Pro.Thissequenceisfoundinhigh
frequencyincollagenandisuniqueamongproteasesinitsabilitytodegradethetriplehelicalnative
collagenfibrilscommonlyfoundinconnectivetissue.
Thecollagenasemostcommonlyusedfortissuedissociationisacrudepreparationcontaining
clostripiopeptidaseAandanumberofotherproteases,polysaccharidasesandlipases.Thiscrude
enzymeisideallysuitedfortissuedissociationsinceitcontainstheenzymerequiredtoattack
nativecollagenandreticularfibers,inadditiontotheenzymeswhichhydrolyzetheotherproteins,
polysaccharidesandlipidsintheextracellularmatrixofconnectiveandepithelialtissues.
Crudecollagenasedoesexhibitlot-to-lotvariabilityandmayproduceoccasionaltoxicity.Invitrogen
attemptedtominimizethesedifficultiesbytissue-typingtheircrudecollagenaselotsbasedupona
correlationbetweenvariousenzymelevelsineachpreparationandeffectivenessindissociating
certaintissues.
Specifications
Potency:Oneunitliberates1μMofL-leucineequivalentsfromcollagenin5hoursat+37°C,
pH7.5.
Types:Particularenzymaticactivitiesofcrudecollagenaseshavecorrelatedwiththe
tissuesfromwhichthecellswereobtained(orwiththeusestowhichthecellsare
put),andasaresultofthecorrelation’s,severalformaltypeshavebeenestablished.
TypeIIISelectedbecauseoflowproteolyticactivity.(caseinassubstrate)
TypeIVSelectedbecauseoflowtrypticactivity.(BAEEassubstrate)
Theseselectedtypeshavebeenfoundtogivebetterperformanceinpreparationofcellsfrom
thevarioustissuesastabulatedbelow.Itshouldbenoted,however,thatwhiletheresults
havebeengreatlyimprovedfollowingthisclassification,thereisstillsomelot-to-lotvariation,
andeffortscontinueinattemptingtogainevenbettercontrolovercrudecollagenase.
TYPETISSUESORCELLS
IFatcells,Adrenal,Liver
IIHeart,Bone,Thyroid,Cartilage,Liver
IIIMammary
IVIslet(insulinreceptorsites)
Inhibitors:Metalchelatingagentssuchascysteine,EDTAoro-phenanthrolinebutnot
DFP.Itisalsoinhibitedbya2-macroglobulin,alargeplasmaglycoprotein.
Instructionsforuse
A.Preparingstockandworkingsolution
Dissolvethenon-sterile,lyophilizedenzymeinHBSS(Cat.No.14025).Filtersterilizethe
solutionwithacellcultureapprovedfiltrationunit.Crudecollagenaseismostoftenused
inconcentrationsfrom0.1to0.5%(W/V)or50to100U/mL.Oncereconstituteduse
immediatelyorstorefrozen.Thawinrefrigeratorimmediatelypriortouse.
B.Dissociationoftissue
•Tissueismincedwithasterilescalpelorscissors.
•WashthetissueseveraltimesinHBSS.
•Thetissuefragmentsaresoakedat+37°C.Increasedefficiencyisobtainedusinga
rockerplatformandsupplementingthedigestwith3mMCaCl2.
C.Organperfusion
•Digestisprewarmedto+37°Candperfusedataratepreoptimizedfortheparticular
organ.Additionof3mMCaCl2increasestheefficiencyofdissociation.
•Dispersedcellsandtissuefragmentsareseparatedfromlargerpiecesbypassingthe
mixturethroughasterilestainlesssteelornylonmesh.Freshcollagenasesolution
canbeaddedtothefragmentsiffurtherdisaggregationisrequired.
•Washseveraltimestoeliminatedebrisandenzymesolution.Adensityseparation
step(Nycodenz)willgiveacleanersuspension.
•Resuspendthepelletintheculturemediumandincubateunderpredetermined
conditions.
ForfurtherinformationonthisorotherGIBCOTMproducts,contactTechnicalServicesatthe
following:
UnitedStatesTECH-LINESM:18009556288
CanadaTECH-LINE:18007578257
OutsidetheU.S.andCanada,refertotheGIBCOproductscataloguefortheTECH-LINEinyour
region.
YoumayalsocontactyourInvitrogenSalesRepresentativeorourWorldWideWebsiteat
www.invitrogen.com.
Forresearchuseonly.
CAUTION:Notintendedforhumanoranimal
diagnosticortherapeuticuses.

胶原酶I	GIBCO[17100-017]	100mg
	GIBCO原装[17100-017]	1g