ProteasomeActivityFluorometricAssayKitIIisdesignedforassayingtheproteasome’schymotrypsin-like,trypsin-likeandcaspase-likeactivitiesinvitrousingpurifiedproteasome,clearedcelllysatesortissueextracts.TheproteasomecleavesthesuppliedfluorogenicsubstratesandthereleasedAMCfluorescencecanbemonitoredusingaplatereaderorfluorometerattheexcitation/emissionwavelengthof360nm/460nm,respectively.
AdditionalInformation
| ProductName: | ProteasomeActivityFluorometricAssayKitII |
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| AlsoKnownAs: | N/A |
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| CatalogNo.: | J4120 |
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| Size: | 1Kit |
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| MolecularWeight: | N/A |
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| Species: | N/A |
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| Source: | N/A |
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| Stock: | 20mMTris,150mMNaCl,2mMβME,10%Glycerol |
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| Concentration: | Seetubelabel |
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| QualityAssurance: | Enzyme>95%bySDS-PAGE;Inhibitorandsubstrate>98%byHPLCandNMR |
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| Storage: | Storeat-80°C;avoidmultiplefreeze-thawcycles |
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| PDFDataSheet: | PDFdatasheet,MSDS |
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| NCBIRefSeq: | N/A |
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| Image(s): | No |
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| ShippingMethod: | Dryiceshipping |
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| References: | N/A |
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Details
*Eachsuppliedfluorogenicsubstrateissufficientfor165x150µlreactions.Thesupplied20Sproteasomeissufficientfor10X150µlreactions.
Description
ProteasomeActivityFluorometricAssayKitIIisdesignedforassayingtheproteasome’schymotrypsin-like,trypsin-likeandcaspase-likeactivitiesinvitrousingpurifiedproteasome,clearedcelllysatesortissueextracts.TheproteasomecleavesthesuppliedfluorogenicsubstratesandthereleasedAMCfluorescencecanbemonitoredusingaplatereaderorfluorometerattheexcitation/emissionwavelengthof360nm/460nm,respectively.Theamountofthecelllysatesortissueextractstobeusedinthereactionshouldbeoptimized.
Suc-LLVY-AMC,Boc-LRR-AMCandZ-LLE-AMCareusedtomonitorthechymotrypsin-like,trypsin-likeandcaspase-likeactivitiesoftheproteasome,respectively.
AMCisusedtoproduceastandardcurveusingaconcentrationrangeof10-100pmol.
Dissolve1mgMG132in105µlDMSOtomakea20mMstocksolutionbeforeuse,using50-200µMMG132toinhibittheproteasomeinvitro.
Thebovine20Sproteasomecanbeusedasapositivecontrol.Atypical150µlreactioncontains10nM20Sproteasome,50µMfluorogenicsubstrateintheassaybuffer.
Components
•AMC(0.1mMinDMSO)....................................................30µl•1000XSUC-LLVY-AMC(50mMinDMSO)........................25µl
•1000XBOC-LRR-AMC(50mMinDMSO)........................25µl•1000XZ-LLE-AMC(50mMinDMSO)...............................25µl
•40XBovine20SProteasome(400nM).............................40µl•MG-132..............................................................................1mg
•DMSO..............................................................................200µl•20XProteasomeAssayBuffer........................................3x1.35ml
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