Skip to main content
. Author manuscript; available in PMC: 2013 Aug 8.
Published in final edited form as: J Biol Chem. 2007 May 11;282(31):22804–22815. doi: 10.1074/jbc.M700961200

FIGURE 1. Purification and fractionation of monoubiquitinated p53 from cells.

FIGURE 1

A, schematic representation of the method for purifying monoubiquitinated p53 from 293 cells. B, protocol for purifying monoubiquitinated p53. C, Western blot analysis of the purified monoubiquitinated p53. Cells were transfected with FLAG-p53, Mdm2, and His-UB and harvested 72 h post-transfection. Western blot analysis after 2 rounds of affinity purification (lane 2) was compared with wild type p53 control (lane 1) using anti-p53 (DO-1 antibody). The migration pattern seen (lane 2) represents monoubiquitination and multi-monoubiquitination.