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. 2009 Jan 30;60(3):993–1008. doi: 10.1093/jxb/ern343

Fig. 9.

Fig. 9.

In-gel CAS activity assay and immunodetection on wild-type OsCAS–YFP homodimer and OsCAS–YFP/OsCAS K99G–HIS heterodimer. In-gel CAS activities on 20 μg of crude proteins from yeast transformants with wild-type OsCAS–YFP (A, lane 1) and co-expressions of OsCAS–YFP/OsCAS K99G–HIS (A, lane 2) were determined. In-gel CAS activity assay on fractions during the purification of the heterodimer through the nickel column from the yeast transformant with co-expressions of OsCAS–YFP/OsCAS K99G–HIS (B) were conducted. Twenty micrograms of fractions from crude extract (lane 1), flow through (lane 2), three washing steps (lanes 3–5), and four elution steps (lanes 6–9) were applied in the assay. Immunodetection of YFP- or HIS-tagged protein on fractions from crude (C, lane 1), flow through (C, lane 2), and purified heterodimer (C, lane 3) were detected by anti-GFP antibody and anti-HIS antibody, respectively.