Figure 2. Western analysis for lipid peroxidation product (4-hydroxynonenal; HNE) modification of isocitrate dehydrogenase. Trabecular meshwork protein extract (5 µg) from each eye with donor age and gender as indicated (M, male; F, female) was loaded on each lane and separated over a 4%–20% SDS–PAGE. The proteins were transferred onto a PVDF membrane and probed using polyclonal antibodies to isocitrate dehydrogenase (isocitrate DH). The corresponding protein bands from the region that showed immunoreactive for isocitrate dehydrgenase was excised from an identical gel and electrophoresed and subsequently transferred on a PVDF membrane and probed for isocitrate dehydrogenase (isocitrate DH) against 4-hydroxynonenal (HNE) and against 3-glyceralde dehydrgenase (GAPDH) as indicated.