Figure 1
Two-step isolation of the mucin protein complexes from human tracheobronchial epithelial (HBE) secretions. An aliquot (10 ml) of HBE secretions was subjected to gel filtration chromatography on an S1000 column. Then, 2-ml fractions were collected and analyzed with periodic acid Schiff (PAS) (pink) and amido black staining (black) (a). The PAS-rich void volume was pooled and subjected to cesium chloride (CsCl) density-gradient centrifugation (b). The fractions were unloaded and probed using slot blotting with PAS (pink) and amido black staining (black) and a keratin sulfate antibody (dotted black). Fractions 1–6, 8–15, and 16–20 were pooled and subjected to proteomic analysis, as reported in Figure 2. The blue line represents density. The intensity units are arbitrary.
