Western blot analysis is shown using Affinity Purified anti-Swi6 antibody to detect endogenous protein present in S.pombe lysate (arrowhead). Comparison to a molecular weight marker (not shown) indicates a band of ~43 kDa corresponding to S.pombe Swi6 protein. ~35ug of lysate was loaded per lane onto a 4-20% gradient gel for SDS-PAGE followed by transfer to 0.45 µm nitrocellulose. The blot was incubated with a 1:1, 700 dilution of the antibody at room temperature for 2 h followed by detection using IRDye (TM)800 labeled Goat-a-Rabbit IgG [H&L] diluted 1:5,000 for 45 min. IRDye (TM)800 fluorescence image was captured using the Odyssey (R) Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.
