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文献和实验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 We describe the facile purification of active E1 from outdated human red blood cells in yields (2–4 nmol/U of blood) that make this an attractive alternative to expression of the proteolytically labile recombinant
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
Isolation of Recombinant DNA Elongation Proteins
This chapter summarizes isolation procedures of four recombinant human proteins crucial for DNA replication: (a) the replicative DNA polymerase (pol) δ, (b) proliferating cell nuclear antigen (PCNA), (c) replication protein A (RP-A), and (d
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