protein

WWE Affinity Resin Set

MBS376021

1 Set

445 USD

Protein

WWE Affinity Resin Set

[WWE]

GST-WWE fusion protein, purity >95% by SDSPAGE

Each vial contains 0.5mg purified GST-WWE fusion protein bound to 50-75ìL packed volume of glutathione beads in 0.5 mL buffer containing 10 mM sodium phosphate, pH 7.4, 150 mM NaCl, 1 mM EDTA, 1% Triton X-100, and 0.02% sodium azide.

Stable for 6 months from date of shipment when stored at 4 degree C. DO NOT FREEZE!

Western Blot (WB), Immunoblot (IB)

Use 20uL (20ug) suspended resin to affinity purify/pull-down poly-ADP-ribose modified proteins in 0.15-1mg cell and tissue extracts. Analyze by Western blotting using protein-specific antibodies to probe the immunoblot. Each 0.5mL vial is sufficient for analysis of ~25 samples. Optimal experimental conditions must be determined by the user.

Testing Data

Additional Notes: The WWE Neg Control Resin has been shown to bind to purified highly automodified PARP1 under certain conditions.

Background: WWE Affinity Resin and Negative Control Resin are designed for the isolation and study of poly-ADP-ribosylated (PARylated) proteins. Through the use of this highly specific PAR affinity resin, PARylated proteins are isolated from cell or tissue lysates without the use of anti-PAR antibodies. The resin-bound proteins can be eluted from the affinity resin, and analyzed by immunoblotting or other methods. RNF146 (Iduna) is a RING-domain E3 ubiquitin ligase that positively regulates Wnt signalling. RNF146 directly interacts with poly(ADP-ribose) through its WWE domain. The WWE domain is a conserved globular domain found in multiple PARPs and E3 ligases. The WWE Affinity Resin is a GST fusion protein of WWE PARbinding domain of human RNF146, amino acid residues 100-175, bound to a glutathione resin. The negative control resin is identical to the WWE Affinity Resin except for a single amino acid substitution that effectively abolishes PAR binding. Description: WWE Affinity Resin is highly purified GST-WWE fusion protein expressed in E. coli, and bound to glutathione beads. The WWE domain sequence is human RNF146 (Iduna) amino acid residues 100-175. It is useful for affinity purification (pulldown) of PARsylated proteins. WWE Negative Control Resin is identical to the WWE Affinity Resin resin except for a single amino acid substitution R163A, which abolishes PAR binding. The negative control resin is useful to control for non-specific binding, and its use is optional.

H.C. Kang et al. (2011) PNAS 108 14103 Y. Zhang et al. (2011) Nature Cell Biol. 13 623 M.G. Callow et al. (2011) PLoS ONE 6 e22595 S.A. Andrabi et al. (2011) Nature Medicine 17 692 Z.D. Zhou et al. (2011) Cell Adh. Migr. 5 463 Z. Wang et al. (2012) Genes & Dev. 26 235

1 Set

445 USD