FIG. 2.

2-D gel analysis of Kin28p. (A) Protein samples containing Kin28p alleles were subjected to isoelectric focusing with a 1:1 mixture of pH 4 to 8 and pH 5 to 7 carrier ampholytes, run on SDS–12.5% polyacrylamide gels in the second dimension, and immunoblotted. To aid in identifying spots, lanes containing Kin28p and Kin28p^T162A samples were run alongside tube gels in the second dimension (a). Kin28p alleles tested included wild-type (WT) Kin28p (b), Kin28p^T162A (c), phosphatase (P’ase)-treated wild-type Kin28p (d), and phosphatase-treated Kin28p^T162A (e). (B) To rule out the possibility of a cross-reacting species in these immunoblots, extracts from cells expressing tagged (a) or untagged (b) Kin28p were subjected to isoelectric focusing with pH 4 to 8 carrier ampholytes and processed as described for panel A.