Human SAE I[1]/SAE II[2] (LAE-E I-SUMO)

(Catalog # P001; P006)

Description:

LAE-E I-SUMO protein is a heterodimer consisted of human SAE I (SUMO-1 activating enzyme subunit 1 [access no: Q9UBE0]) and SAE II (Ubiquitin-like protein activating enzyme [access no: Q9UBT2]). Both the SAE I and SAE II are co-expressed in a baculovirus expression system, prepared from freshly extracted cell lysates, and purified to homogeneity using methods developed by LAE Biotechnology Co., LTD. The final fraction of enzyme contains two single polypeptide bands of 38 kDa and 80 kDa[3].

Storage Buffer:

The final fraction of LAE-E I-SUMO 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 1mM DTT, and 10 % glycerol. The enzyme is stable in this buffer at -70^oC. In general, the enzyme is less stable when diluted. We do not  recommend any dilution of the enzyme. In addition, it is a good practice to aliquot the enzyme after the first thaw .

Concentration:

LAE-E I-SUMO is supplied at the concentration of 75 ng/ul (Cat # P001: 15 ug/tube, 200ul/tube; Cat # P006: 7.5ug/tube, 100 ul/tube).

Assay Conditions:

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

Shipping/Storage:

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

Legal consideration: FOR RESEARCH USE ONLY.

Left: Human topo I (S³⁵Met-labeled) control.

Right: Sumoylated human topo I (S³⁵Met-labeled)

(Sumoylation was performed at 37 ^oC for 30 min in the presence of 150 ng of E1, 1 ug of E2, and 1 ug of sumo1in a final volume of 20 ul with reaction buffer 20 mM Hepes pH 7.5, 5 mM MgCl₂, and 2 mM ATP).