Nedd8ConjugationKitisdesignedforassayingtheconjugationofNedd8toproteinsubstratesinvitro.Inaproteinneddylationreaction,Nedd8isfirstactivatedbytheNedd8activatingenzymeAPPBP1/UBA3(E1)inanATPhydrolysis-dependentmanner;activatedNedd8isthentransferredtotheNedd8conjugatingenzymeUbe2M(UbcH12)(E2)byformationofathiolesterbondwiththecatalyticcysteineresidueofUbcH12I;Nedd8-chargedUbe2M(UbcH12)canmediateproteinneddylationinthepresenceofanappropriateNedd8ligase(E3).
AdditionalInformation
| ProductName: | Nedd8ConjugationKit |
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| AlsoKnownAs: | N/A |
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| CatalogNo.: | J3150 |
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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): | No |
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| ShippingMethod: | Dryiceshipping |
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| References: | N/A |
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Details
*Thiskitissufficientforsettingup25x20µlreactions.
Description
Nedd8ConjugationKitisdesignedforassayingtheconjugationofNedd8toproteinsubstratesinvitro.Inaproteinneddylationreaction,Nedd8isfirstactivatedbytheNedd8activatingenzymeAPPBP1/UBA3(E1)inanATPhydrolysis-dependentmanner;activatedNedd8isthentransferredtotheNedd8conjugatingenzymeUbe2M(UbcH12)(E2)byformationofathiolesterbondwiththecatalyticcysteineresidueofUbcH12I;Nedd8-chargedUbe2M(UbcH12)canmediateproteinneddylationinthepresenceofanappropriateNedd8ligase(E3).
Atypical20µlreactioncontains100nMAPPBP1/6xHis-UBAs,2µM6xHis-Ube2M(UbcH12),15µM6xHis-Nedd8,4mMATP,2µME3(notsupplied)and2µMproteinsubstrate(nosupplied)intheNeddylationBuffer.
Components
•20XAPPBP1/6xHis-UBA3(2µM)................25µl•20X6xHis-Ube2M(UbcH12)(40µM))..........25µl
•10X6xHis-Nedd8(150µM)..........................50µl•10XATP(40mM)........................................60µl
•10XNeddylationBuffer...............................100µl
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