ProductDescription
Collagenaseconsistsofablendofenzymesincludingcollagenase,caseinase,clostripainandtrypsindesignedtoeffectivelyhydrolyzecollagenanddissociatetissues.VariouscollagenaseproductshavebeenevaluatedspecificallyforusewithAdvancedBiomatrix’scollagenproducts.ThisproductisofferedtoimprovetheviABIlityandfunctionalityofisolatedcellsfromthedigestedcollagengels.
Collagenaseisprovidedasalyophilized,sterilepowderina50mgpackagesize.Afterreconstitution,theproductisready-to-use.
| Parameter,Testing,andMethod | Collagenase#5030 |
| Form | LyophilizedPowder |
| PackageSize | 50mg |
| StorageTemperature | 2-10°C |
| SterilizationMethod | Filtration |
| Bioburden(cfu) | <1 |
| Source | ClostridiumHistolyticum |
| ShelfLife | Minimumof6monthsfromdateofreceipt |
DirectionsforUse
DownloadtheDirectionsforUsePDForcontinuereADIngbelow:
- Usetheserecommendationsasguidelinestodeterminetheoptimalgeldissociationprocedureforyourculturesystem.
- Reconstitutethelyophilizedcollagenasepowder(50mg)inphosphatebufferedsalineorotherpHneutralmedium.Atypicalconcentrationof1mg/mlmaybeused.
- Aspiratetheculturemediumfromthecollagengel.
- Washthecollagengel2Xwithpre-warmedPBSorotherpHneutralmediumtoremoveculturemediumwhichcouldinhibitthedegradationofcollagen.
- Addappropriateamountofcollagenasesolution(onegelvolumeshouldbesufficient).Forexample:For24wellplates,containing0.5mlofgel,add0.5mlofcollagenasesolution.
- Transfertoa37ºCincubatorfor30-60minutes.Tofacilitatethefasterdissociationofthecollagengel,thegelandcollagenasesolutioncanbePipettedupanddownusingalargeborepipettetip.
- Incubateadditional30-60minutesat37°C,ifnecessary,tocompletethedigestionofthegel.Ifrequired,pipettethegel/cellmixtureevery15minutes.Note:Thickergelsorgelscontaininghigherconcentrationofcellsmayrequiremoretime.
- Oncethegelisfullydigested,addanequalvolumeofcompleteculturemediumtothegel/cellmixtureandthencollectthecells.
- Rinsetheculturevesselcollectinganyresidualcells.
- CentrifugethecellsUSPensionat250xgfor5minutesatroomtemperature.
- Carefullyaspiratethesupernatant,andresuspendthecellpelletin0.5-1mloffreshmedium.
- DeterminecellcountandviabilityusingahemocytometerandTrypanBlue.
ProductQ&A
Yes.
Therearesufficientlevelsofdivalentcationsinthelyophilizedcollagenasepowdertoachievefullenzymaticactivity.Therefore,solubilizingthelyophilizedcollagenpowderinmediumoraphosphatebufferedsolutionwithoutdivalentcationsisrecommendedpertheDirectionsforUse.
Collagenaseismoststableinthelyophilizedformstoredat2to10°C.Afterreconstitution,itisrecommendedthatanyremainingsolubilizedcollagenasebealiquotedintodesiredvolumesandthenfrozenat<-20°Cforupto1to2months.Repeatfreezingandthawingisnotrecommended.
ProductReferences
CollagenaseReferences:
Röck,Katharina,etal."CollagenfragmentsinhibithyaluronansynthesisinskinfibroblastsinresponsetoultravioletB(UVB)newinsightsintomechanismsofmatrixremodeling."JournalofBIOLOGicalChemistry286.20(2011):18268-18276.
Shieh,HesterF.,etal."ComparisonsofhumanamnioticmesenchymalstemcellviabilityinFDA-approvedcollagen-basedscaffolds:Implicationsforengineereddiaphragmaticreplacement."Journalofpediatricsurgery52.6(2017):1010-1013.
Kumar,BalasubramanianK.,etal."Theeffectsofoverexpressingaquaporinsonthecryopreservationofhepatocytes."Cryobiology71.2(2015):273-278.
Yang,Xi,andWilliamF.Salminen."Kavaextract,anherbalalternativeforanxietyrelief,potentiatesacetaminophen-inducedcytotoxicityinrathepaticcells."Phytomedicine18.7(2011):592-600.
Ingeson-Carlsson,Camilla,AngelaMartinez-Monleon,andMikaelNilsson."DifferentialeffectsofMAPKpathwayinhibitorsonmigrationandinvasivenessofBRAFV600Emutantthyroidcancercellsin2Dand3Dculture."Experimentalcellresearch338.2(2015):127-135.
Voedisch,Sabrina,etal."Neuropeptidescontrolthedynamicbehaviorofairwaymucosaldendriticcells."PloSone7.9(2012):e45951.
ProductCertificateofAnalysis
SafetyandDocumentation
SafetyDataSheet
CertificateofOrigin
ProductDisclaimer
ThisproductisforR&Duseonlyandisnotintendedforhumanorotheruses.PleaseconsulttheMaterialSafetyDataSheetforinformationregardinghazardsandsafehandlingpractices.
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