K-220

Material Data Sheet

RNF4 Ubiquitin Ligase Kit - di-SUMO3 Substrate

Price: $325.00
Catalog #: K-220

RNF4 Ubiquitin Ligase Kit - di-SUMO3 Substrate

Price: 
$325
Quantity: 
1 Kit
Data Sheet: 

RNF4 (SNURF) is a RING-finger ubiquitin E3 ligase that ubiquitinates and mediates the proteasomal destruction of PML/RARα, a fusion protein that is a hallmark of acute promyelocytic leukemia (APL). APL can sometimes be treated effectively with arsenic trioxide, which induces PML/RARα modification by small ubiquitin-like modifiers (SUMO). RNF4 subsequently binds to and ubiquitinates the poly-SUMOylated PML/RARα, thereby targeting it to the proteasome.

RNF4 contains four SUMO-interacting motifs (SIMs) that function to recruit this ligase to a variety of poly-sumoylated substrates. RNF4 will autoubiquitinate in vitro, and will also ubiquitinate poly-SUMO chains. This kit is designed for in vitro RNF4-mediated ubiquitination of user-supplied, poly-SUMOylated substrates such as PML, PEA3, CENP1, and PARP1. The resulting proteins can be used in downstream applications, or analyzed by Western blot using antibodies specific for the target protein. A control substrate, di-SUMO3 (ULC-300) and detection antibody (MAB2959) are included in the kit. 

NOTE: Kit contains reagents sufficient for 10 x 30 μl reactions and 5 Western Blots (mini-gel format).

Storage: 

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

Literature

References: 

Geoffroy, M-C., et al. (2010)  Mol. Bio. Cell   21:  4227-4239

Hakli M., et al.  (2004)  FEBS Lett.   560: 56-62

Tatham M.H., et al.  (2008)  Nat. Cell Biol.  10:  538-546

Vertegaal A.C.O.  (2011)  Chem. Rev111: 7923–7940