K-210
Mdm2/HDM2 Ubiquitin Ligase Kit - S5a Substrate
Mdm2/HDM2 Ubiquitin Ligase Kit - S5a Substrate
The RING-finger ubiquitin E3 ligase Mdm2/HDM2 (Murine Double Minute 2) is an important regulator of the tumor suppressor protein p53. Mdm2/HDM2 ligase activity is at least partially responsible for the ubiquitination and subsequent proteasomal degradation of p53 and other proteins. This kit is designed for in vitro Mdm2/HDM2-mediated ubiquitination of user-supplied substrates such as p53 (SP-450), 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.
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
Brooks C.L., et al. (2007) J.Biol.Chem 282: 22804-22815
Bushmann T., et al. (2001) J.Biol.Chem 276: 40389-40394
Dornan D, et al. (2004) Nature 429:86-92
Grossman S.R., et al. (2003) Science 300:342-344
Hu M., et al (2006) PLoS Biol. 4:228-239
Kawai H., et al. (2003) Mol. Cell. Biol. 23: 4939-4947
Lai Z., et al. (2001) J.Biol.Chem 276: 31357-31367
Li M, et al. (2003) Science 301:1972-1975
Linares L.K., et al. (2003) Proc.Natl.Acad.Sci. 100:12009-12014
Meulmeester E. and Jochemsen A.G. (2008) Curr.Canc.Drug. Targets 8:87-97
Sheng Y, et al. (2006) Nat.Struc.Mol.Biol. 13:285-291
Wu H., et al. (2011) Nature Med. 17: 347-356
Yan Y. and Chen J. (2005) Biochem. Biophys. Res. Comm. 332:702-709
Yu G.W., et al. (2006) Proc.Natl.Acad.Sci. 103:1227-1323