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. 2009 Mar 5;106(12):4584–4590. doi: 10.1073/pnas.0900774106

Fig. 4.

Fig. 4.

Separation of tryptic phosphopeptides from Pellino 1. 32P-labeled Pellino 1 obtained by incubation with Mg-[γ32P]ATP and either IRAK4 (A and B) or IRAK1 (C) was subjected to SDS/PAGE, the gel was stained with Coomassie blue, and Pellino 1 was excised, digested with trypsin, and subjected to RP-HPLC on a Vydac C18 column equilibrated in 0.1% (vol/vol) trifluoroacetic acid. The column was developed with an acetonitrile gradient in 0.1% (vol/vol) trifluoroacetic acid. 32P radioactivity in arbitrary units (au) is shown by the full line, and the acetonitrile gradient is indicated by the diagonal broken lines. (A and C) Wild-type Pellino 1. (B) Pellino 1[8A].