FIG. 3.

Immunoblotting of B. burgdorferi membrane-associated proteins separated by 2DE with Lyme patient sera. MAP were separated with pH 4-7 2DE-IPG (A to C) or 2DE-NEPHGE (D to F) and immunoblotted with a 1:1,000 dilution of pooled CDC Lyme patient sera representing early-localized (A and D), early-disseminated (B and E), and late-disseminated (C and F) stages of Lyme disease. The pH range of each gel is indicated. Relative molecular mass markers (in kilodaltons) are indicated to the left of each gel. Proteins identified by MALDI-TOF analysis of identical silver-stained spots are annotated. Boxed areas designate protein isoforms with a single identification. For pH 4-7 2DE-IPG, 400 μg of MAP was used for silver staining and 50 μg of MAP was used for immunoblotting. 2DE-NEPHGE utilized 150 μg of MAP for silver staining and 25 μg of MAP for immunoblotting.