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上海玉博生物科技有限公司
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文献和实验Performing In Vitro Sumoylation Reactions Using Recombinant Enzymes
to perform in vitro sumoylation reactions, namely the E1 activating enzyme Aos1/Uba2 (SAE1/SAE2), the E2 conjugating enzyme Ubc9, SUMO-1 (identical protocols can be used for SUMO-2/3), and the catalytic domain of the E3 ligase RanBP2/Nup358. Two alternative
Purification of E1 and E1-Like Enzymes
that has been adapted for the isolation of E1 paralogs. We describe the facile purification of active E1 from outdated human red blood cells in yields (2–4 nmol/U of blood) that make this an attractive alternative to expression of the proteolytically labile recombinant
An In Vitro FRET-Based Assay for the Analysis of SUMO Conjugation and Isopeptidase Cleavage
and purifi ed YFP-SUMO-1 and CFP-RanGAP1 as model substrates that are covalently coupled in the presence of recombinant SUMO E1 and E2 enzymes and ATP. Reactions of 25 μl volume, set up in 384-wells plates, give suffi cient signal for analysis. Consequently
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