Human Aos1/Uba2 (E I-SUMO)

(Catalog # P006)

Description:

Human Aos1/Uba2 protein is a heterodimer consisted of human AOS1 ( [access no: NP_00549]; also called SUMO-1 activating enzyme subunit 1; SUMO-1 activating enzyme E1 N subunit; sentrin/SUMO-activating protein AOS1; ubiquitin-like protein SUMO-1 activating enzyme) and Uba II ([access no: AAD23914]; also called hUBA2; ubiquitin-like protein activating enzyme; sentrin activating enzyme). Both the Aos I and Uba II are co-expressed in a baculovirus expression system, prepared from freshly extracted cell lysates, and purified to homogeneity using methods developed by Corgen Inc. The final fraction of enzyme contains two single polypeptide bands of 38 kDa and 80kDa.

Storage Buffer:

The final fraction of Human Aos1/Uba2 protein is an affinity column pool and is in the following buffer: 10 mM Tris-Cl, pH 7.5, 100 mM NaCl, 100 mM imidazole, 0.5 mM PMSF, 1 mM DTT, and 50 % glycerol. The enzyme is stable in this buffer at -20ºC. In general, the enzyme is less stable when diluted. We don’t recommend any dilution of the enzyme. In addition, it is a good practice to aliquot the enzyme after the first thaw.

Concentration:

Human Aos1/Uba2 protein is supplied at the concentration of 0.15 µg/µl (Cat # P006: 4.5µg/tube, 30µl/tube).

Assay Conditions:

Sumoylation assays are carried out in a final volume of 20 µl in reaction buffer (20 mM Hepes pH 7.5, 5mM MgCl2, 2mM ATP). We would recommend to add fresh 2mM ATP to be sure that sufficient energy is supplied.

Shipping/Storage:

The enzyme is shipped on dry ice and must be stored at -70ºC or lower.

Legal consideration: FOR RESEARCH USE ONLY.

Reference:

Desterro,J.M., Rodriguez,M.S., Kemp,G.D. and Hay,R.T., Identification of the enzyme required for activation of the small ubiquitin-like protein SUMO-1., J. Biol. Chem. 274 (15), 10618-10624 (1999)  MEDLINE   99214633   PUBMED   10187858
　

Gong,L., Li,B., Millas,S. and Yeh,E.T., Molecular cloning and characterization of human AOS1 and UBA2, components of the sentrin-activating enzyme complex., FEBS Lett. 448 (1), 185-189 (1999)  MEDLINE   99231789   PUBMED   10217437
　

Okuma,T., Honda,R., Ichikawa,G., Tsumagari,N. and Yasuda,H., In vitro SUMO-1 modification requires two enzymatic steps, E1 and E2., Biochem. Biophys. Res. Commun. 254 (3), 693-698 (1999)  MEDLINE   99120925   PUBMED   9920803
　

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