Sumo1,Sumo2,Sumo3ConjugationKitisdesignedforassayingtheconjugationofSumo1,Sumo2andSumo3toproteinsubstratesinvitro.Inaproteinsumoylationreaction,aSumoproteinisfirstactivatedbytheSumoactivatingenzymeSAE1/SAE2(E1)inanATPhydrolysis-dependentmanner;activatedSumoisthentransferredtotheSumoconjugatingenzymeUbe2I(E2)byformationofathiolesterbondwiththecatalyticcysteineresidueofUbe2I;Sumo-chargedUbe2IcanmediateproteinsumoylationinthepresenceofanappropriateSumoligase(E3).
AdditionalInformation
| ProductName: | Sumo1,Sumo2,Sumo3ConjugationKit |
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| AlsoKnownAs: | N/A |
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| CatalogNo.: | J3140 |
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| Size: | 1Kit |
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| MolecularWeight: | N/A |
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| Species: | Human |
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| Source: | Bacterialrecombinant |
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| Stock: | 20mMTris,150mMNaCl,2mMβME,10%Glycerol |
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| Concentration: | Seetubelabel |
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| QualityAssurance: | 95%basedonSDS-PAGE |
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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): | N/A |
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| ShippingMethod: | Dryiceshipping |
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| References: | N/A |
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Details
*Thiskitissufficientforsettingup25x20µlreactions.
Description
Sumo1,Sumo2,Sumo3ConjugationKitisdesignedforassayingtheconjugationofSumo1,Sumo2andSumo3toproteinsubstratesinvitro.Inaproteinsumoylationreaction,aSumoproteinisfirstactivatedbytheSumoactivatingenzymeSAE1/SAE2(E1)inanATPhydrolysis-dependentmanner;activatedSumoisthentransferredtotheSumoconjugatingenzymeUbe2I(E2)byformationofathiolesterbondwiththecatalyticcysteineresidueofUbe2I;Sumo-chargedUbe2IcanmediateproteinsumoylationinthepresenceofanappropriateSumoligase(E3).
Atypical20µlreactioncontains100nMSAE1/SAE2,2µM6xHis-Ube2I,50µMSumo1(orSumo2,orSumo3),4mMATP,2µME3(notsupplied)and2µMproteinsubstrate(nosupplied)inthesumoylationbuffer.
Components
•20XSAE1/SAE2(2µM).........................25µl•20X6xHis-Ube2I(40µM)........................25µl
•10XSumo1(500µM)..............................50µl•10XSumo2(500µM)..............................50µl
•10XSumo3(500µM)..............................50µl•10XATP(40mM).................................125µl
•10XSumoylationBuffer...........................1ml
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