Fig 2.

1-DZ analysis and identification of gelatinases by Western blot. 1.5 μl of serum from healthy control (HC) or from a patient with inactive RR-MS were loaded on a 7.5% polyacrylamide gel copolymerized with 0.1% (w/v) gelatin (1-DZ). For Western blot analysis, 2 μl of fivefold concentrated (Vivaspin 500, MWCO 50,000; GE Healthcare) serum samples (the same as in 1-DZ) were separated on a 7.5% polyacrylamide gel without gelatin (1-DE), transferred on nitrocellulose membranes and incubated with the following primary antibodies: Anti-NGAL Antibody (5G5) at concentration of 2.0 μg/ml; Anti-MMP-9 (Ab-8) Mouse mAb (IA5) at concentration of 2.66 μg/ml; Anti-MMP-2 (Ab-4) Mouse mAb (75-7F7) at concentration of 1.0 μg/ml and Anti-MMP-9 Ab (4A3) at concentration of 2.0 μg/ml. Standard Proteins (SP): Precision Plus Protein Standards Dual Color (Bio-Rad).