Affinity-based assays. (A) Western blot of total lens extract, probed with rabbit antibody to the recombinant, purified ppl-L, characterizing the affinity-purified antibody. (B) Total lens detergent extracts of wild-type lenses (lanes 1, 2) and vimentin knockout lenses (lanes 3, 4). Lanes 1 and 3 are controls probed with irrelevant rabbit IgG. Lanes 2 and 4 are probed with the antibodies indicated at the left.(C) GST pull-down assays. Immobilized GST-ppl-L fusion protein was used to probe detergent-extracted lens. The washed pellet was resolved by SDS-PAGE and transferred to nitrocellulose for probing with antibodies to (lane 1) CP49, (lane 2) filensin, (lane 3) vimentin.