Fig 5.

Identification of gelatinases by Western blot analysis. A pool of sera from 10 patients with inactive RR-MS was used. For 2-DZ, pooled sera were concentrated 10-fold and 40 μg of total proteins were applied (left panels). For Western blot analysis pooled sera were concentrated 40-fold and 160 μg of total proteins were subjected to 2-DE (right panels). 2-DE gels were cut and the corresponding nitrocellulose membranes were treated with anti-NGAL Ab (A'), anti-MMP-2 Ab (B') and Anti-MMP-9 (4A3) Ab, recognizing the 65 kD and the 82 kD activated forms (C'). Gel A shows the region with isoelectric point between pH 5–6.5 and molecular mass between 200 and 75 kD, analysed by Western blot in A'; Gels B and C show the region with isoelectric point between pH 4–5.5 and molecular mass between 85 and 55 kD, analysed by Western blot in B' and C', respectively.