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.