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. 2000 Nov;157(5):1439–1445. doi: 10.1016/S0002-9440(10)64781-5

Figure 2.

Figure 2.

Western blot analysis of 3-NT pAb antibody. A: Coomassie-stained gel of in vitro nitrated α-syn, tubulin, NFL, NFM, and NFH. Five μg of each protein were resolved by electrophoresis on separate lanes of a discontinuous 6% to 10% to 12% sodium dodecyl sulfate-polyacrylamide gel. The arrow on the left denotes monomeric α-syn. B: In Western blots the 3-NT pAb variably recognizes each of the nitrated proteins but none are as intensely immunoreactive as nitrated α-syn. Note the robust recognition of monomeric α-syn as well as higher oligomeric α-syn species by this antibody. The arrows in B identify the far less intensely labeled immunobands for each of the other proteins. Fifty ng of each protein were resolved by electrophoresis on a discontinuous 6% to 10% to 12% sodium dodecyl sulfate-polyacrylamide gel. After electrophoretic transfer to nitrocellulose membrane, the blot was blocked with 5% skimmed milk, sequentially incubated with the 3-NT pAb and anti-rabbit-horseradish peroxidase-conjugated antibody and developed with enhanced luminol reagents (Dupont-New England Nuclear). The position of the molecular weight markers are indicated on the left.