Validating PPAR Antibody Specificity

With available PPARα knockout tissue, we tested the specificity of the PPARα antibody. As shown in Supplemental Fig. S1A, there was no immunoreactivity of the PPARα antibody in a PPARα knockout—confirming specificity of this antibody. As for PPARγ, Sarruf et al. tested five commercially available PPARγ antibodies (including the Abcam rabbit anti-PPARγ used in this study) and found similar immunoreactivity patterns and a reduction in signal in floxed PPARγ knockouts using the antibody used in this study46 (link)47 (link). If knockout tissue is not easily obtainable or previously published, one can also use immunoblots to determine whether the primary antibody can bind to a single protein of the correct molecular weight. For PPARβ/δ other researchers have confirmed the specificity of the primary antibody used in this study using western blots15 (link)48 (link).