K-230

Material Data Sheet

E6AP Ubiquitin Ligase Kit - S5a Substrate

Price: $395.00
Catalog #: K-230

E6AP Ubiquitin Ligase Kit - S5a Substrate

Price: 
$395
Quantity: 
1 Kit
Data Sheet: 

E6AP (E6-Associated Protein) is a HECT domain ubiquitin E3 ligase that ubiquitinates and mediates the proteasomal destruction of substrate proteins.  HECT domain ligases use an active site cysteine to accept charged ubiquitin from ubiquitin-E2 thioester complexes for subsequent transfer to substrate proteins; in this way HECT class ligases are distinct from most RING class ligases in that the latter facilitate transfer of ubiquitin from charged E2’s directly to substrate proteins without an E3-ubiquitin thioester intermediate.  This kit is designed for in vitro E6AP-mediated ubiquitination of user-supplied, reported substrates such as β-Arrestin, and DYRK2.  Ubiquitinated proteins can be used in downstream applications, or analyzed by Western blot using antibodies specific for the target protein.  A control substrate, S5a (SP-400) and detection antibody (AF5540) are included in the kit.

Storage: 

Store protein components at -80oC. Avoid multiple freeze/thaw cycles. Loading Buffer may be stored at room temperature. Mg2+-ATP and α-S5a antibody may be stored at -20oC.

Literature

References: 

Huang L., et al.  (1999)  Science   286: 1321-1326

Kim H.T. & Goldberg A.L. (2012)  Methods in Mol. Bio.  832: 653-660

Martinez-Noel G., et al.   (2012)   Mol. Cell. Biol.   32: 3095-3106

Mishra A., et al.  (2009)   J Biol Chem.  284: 10537-10545