Figure 2

Wld^S mice can be accurately genotyped by quantitative western blots of tail tip protein expression. Tail tips were homogenized in RIPA buffer and following a BCA assay, 30 μg of protein were loaded per lane of a pre-cast gradient gel (see methods section). A - Representative example bands obtained with quantitative fluorescent western blots from a litter of mice produced by a heterozygous breeding pair. The litter contained wild-type, heterozygous and homozygous Wld^Smice. Blots were probed for Wld^S protein levels using the Wld18 antibody. Tubulin was used as a loading control (data not shown). B - Wld^S protein expression shown as arbitrary fluorescence units, illustrating that protein expression was roughly doubled in homozygous mice compared to heterozygous mice. N = 4 wild-type mice, N = 3 heterozygous Wld^Smice and N = 5 homozygous Wld^Smice.