Sumo1 Conjugation Kit

Sumo1 Conjugation Kit is designed for assaying the conjugation of Sumo1 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).N/A

Additional Information

Product Name: Sumo1 Conjugation Kit
Also Known As: N/A
Catalog No.: J3110
Size: 1 Kit
Molecular Weight: N/A
Species: Human
Source: Bacterial recombinant
Stock: 20 mM Tris, 150 mM NaCl, 2 mM βME, 10% Glycerol
Concentration: See tube label
Quality Assurance: 90% - 95% based on SDS-PAGE
Storage: Store at -80°C; avoid multiple freeze-thaw cycles
PDF Data Sheet: PDF datasheet, MSDS
NCBI RefSeq: N/A
Image(s): No
Shipping Method: Dry ice shipping
References: N/A

Details

* This kit is sufficient for setting up 25 x 20 µl reactions.

Description

Sumo1 Conjugation Kit is designed for assaying the conjugation of Sumo1 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, 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 ATP (40 mM)....................................60 µl
• 10X Sumoylation Buffer.........................100 µl