FIG. 3.

Affinity blotting of cell-derived proteins, using representative recombinant ScaD cohesins as probes. A. cellulolyticus cells were grown on cellobiose and pelleted, and the supernatant fraction was adsorbed onto phosphoric acid-treated cellulose. Samples were subjected to SDS-PAGE (Gel), and blotted onto nitrocellulose membranes (Blots). Gels were stained with Coomassie brilliant blue. The blots were probed with the designated recombinant protein sample, and labeled bands were detected by chemiluminescence with peroxidase-conjugated, anti-His tag antibody. Probes: CohD1, the first cohesin of ScaD; CohD3, the third cohesin of ScaD. Std, prestained molecular weight markers (in thousands). Sup, original sample of extracellular protein, adsorbed onto cellulose.