Sumo1, Sumo2, Sumo 3 Conjugation Kit is designed for assaying the conjugation of Sumo1, Sumo2 and Sumo3 to protein substrates in vitro. In a protein sumoylation reaction, a Sumo protein is first activated by the Sumo activating enzyme SAE1/SAE2 (E1) in an ATP hydrolysis-dependent manner; activated Sumo is then transferred to the Sumo conjugating enzyme Ube2I (E2) by formation of a thiolester bond with the catalytic cysteine residue of Ube2I; Sumo-charged Ube2I can mediate protein sumoylation in the presence of an appropriate Sumo ligase (E3).

Additional Information

Product Name: Also Known As: Catalog No.: Size: Molecular Weight: Species: Source: Stock: Concentration: Quality Assurance: Storage: PDF Data Sheet: NCBI RefSeq: Image(s): Shipping Method: References:

Sumo1, Sumo2, Sumo 3 Conjugation Kit

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J3140

1 Kit

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Human

Bacterial recombinant

20 mM Tris, 150 mM NaCl, 2 mM βME, 10% Glycerol

See tube label

95% based on SDS-PAGE

Store at -80°C; avoid multiple freeze-thaw cycles

PDF datasheet, MSDS

N/A

N/A

Dry ice shipping

N/A

Details

* This kit is sufficient for setting up 25 x 20 µl reactions.DescriptionSumo1, Sumo2, Sumo 3 Conjugation Kit is designed for assaying the conjugation of Sumo1, Sumo2 and Sumo3 to protein substrates in vitro. In a protein sumoylation reaction, a Sumo protein is first activated by the Sumo activating enzyme SAE1/SAE2 (E1) in an ATP hydrolysis-dependent manner; activated Sumo is then transferred to the Sumo conjugating enzyme Ube2I (E2) by formation of a thiolester bond with the catalytic cysteine residue of Ube2I; Sumo-charged Ube2I can mediate protein sumoylation in the presence of an appropriate Sumo ligase (E3).A typical 20 µl reaction contains 100 nM SAE1/SAE2, 2 µM 6xHis-Ube2I, 50 µM Sumo1 (or Sumo2, or Sumo3), 4 mM ATP, 2 µM E3 (not supplied) and 2 µM protein substrate (no supplied) in the sumoylation buffer.Components• 20X SAE1/SAE2 (2 µM).........................25 µl• 20X 6xHis-Ube2I (40 µM)........................25 µl• 10X Sumo1 (500 µM)..............................50 µl• 10X Sumo2 (500 µM)..............................50 µl• 10X Sumo3 (500 µM)..............................50 µl• 10X ATP (40 mM).................................125 µl• 10X Sumoylation Buffer...........................1 ml