Figure 6

Electrospun membranes in Western blotting applications. (A). Chemiluminescence detection of Fn (lanes 1 to 9 = 10.0, 5.0, 2.5, 1.0, 0.50, 0.25, 0.16 0.08, 0.02 μg Fn/lane) on parent nitrocellulose and electrospun nitrocellulose, parent nylon and electrospun nylon (using 1 μm diameter fibers). Electrospun nitrocellulose exhibited high signal, but poor band resolution. The parent nylon material exhibited a diffuse signal in lanes loaded with the highest concentration of Fn and prominent negative images (A, lanes 1–5, top right). Contrast with electrospun nylon, this membrane exhibited sharper bands and no evidence of inverse image formation. (B) Electrospun nylon/electrospun nitrocellulose fiber composite. Note: high signal, absence of inverse white bands, but poor band resolution. We associate this result with a composite that is too thick. SEM images before (Bar = 10 μm) and after (Bar = 5 μm) blocking buffers applied to composite, inset demonstrating material coating fibers, pores between adjacent fibers remain present and open. These images suggested that electrospun nitrocellulose fibers underwent an increase in diameter during blotting (C). Electrospun nylon/electrospray nitrocellulose bead composite. This composite provided good signal detection and band resolution. SEM images before (Bar = 10 μm) and after (Bar = 5 μm) blocking buffers applied to composite.