N-terminal Biotin-Ubiquitin

A single biotin moiety is conjugated on the N-terminus of ubiquitin. All lysine residues of ubiquitin are still available for formation of polyubiquitin chains. The extremely high binding affinity between biotin and streptavidin allows the use of N-terminal biotin-ubiquitin for the following in vitro assays: 1) rapid and efficient purification of polyubiquitinated substrate proteins; 2) monitoring substrate ubiquitination using an HRP-conjugated streptavidin antibody; and 3) setting up high throughput FRET assays to monitor substrate ubiquitination in assays screening E3 ubiquitin ligase inhibitors.

Additional Information

Product Name: N-terminal Biotin-Ubiquitin
Also Known As: Biotin-Ub
Catalog No.: E1140
Size: 50 µg
Molecular Weight: 8.9 kDa
Species: Human
Source: Synthetic
Stock: 20 mM Tris, pH 7.6 at 4 °C, 100 mM NaCl, 10% glycerol
Concentration: 1 mg/ml in aqueous buffer
Quality Assurance: > 95% by HPLC
Storage: Store at -80°C; avoid multiple freeze-thaw cycles
PDF Data Sheet: PDF DatasheetMSDS
NCBI RefSeq: N/A
Image(s): (Click image to enlarge)

LC-MS of N-terminal Biotin-Ubiquitin
Shipping Method: Dry ice shipping
References: N/A

Details

A single biotin moiety is conjugated on the N-terminus of ubiquitin. All lysine residues of ubiquitin are still available for formation of polyubiquitin chains. The extremely high binding affinity between biotin and streptavidin allows the use of N-terminal biotin-ubiquitin for the following in vitro assays: 1) rapid and efficient purification of polyubiquitinated substrate proteins; 2) monitoring substrate ubiquitination using an HRP-conjugated streptavidin antibody; and 3) setting up high throughput FRET assays to monitor substrate ubiquitination in assays screening E3 ubiquitin ligase inhibitors.

Use
Depending on ubiquitination assay conditions, N-terminal biotin-ubiquitin can used in conjunction with wild type ubiquitin at combined concentration range of 20-50 μM with a 1:4 to 1:20 ratio of N-terminal biotin-Ub: wild type ubiquitin.

Images:
(Click image to enlarge)

LC-MS of N-terminal Biotin-Ubiquitin