Skip to main content
. Author manuscript; available in PMC: 2011 Apr 1.
Published in final edited form as: J Neurochem. 2010 Jan 20;113(2):402–417. doi: 10.1111/j.1471-4159.2010.06605.x

Figure 5.

Figure 5

Abolishment of the PRK8 and PRK28 antibody epitopes due to the E399Q human parkin mutant. A) pDEST15/parkin 380–465 and pDEST15/parkin 380–465 E399Q constructs were used to express the respective proteins in BL21 E. coli. Total protein lysates were extracted and equal amounts of protein (600 ng) were resolved on 13% polyacrylamide gels and analyzed by western blot with GST, PRK8, PRK28, and PRK109 antibodies. B) Full-length untagged wild-type and mutant (T240R and E399Q) human parkin cDNA cloned in the mammalian express vector pcDNA3.1 were used to express these proteins in mouse N2A neuroblastoma cells. Following transfection with the respective constructs, equal amounts (4 ug) of total protein lysates were resolved on 13% polyacrylamide gels and assessed by western blot for recognition of the indicated parkin antibodies. Additionally, the blots were probed with an actin antibody to confirm equal protein loading. The mobility of molecular mass markers is indicated on the left.