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. 2009 Oct;128(2):206–217. doi: 10.1111/j.1365-2567.2009.03101.x

Figure 5.

Figure 5

Substrate-dependent and -independent ubiquitylation. (a) Ubiquitin ligase assays (16 hr) were performed as described in the Materials and methods using recombinase activating gene 1 (RAG1)[218–389], CDC34 and RAG1[264–389] as indicated. Products were analysed by western blot with anti-ubiquitin conjugate antibody followed by anti-rabbit-horseradish peroxidase (HRP) antibody. (b) Staging scheme for reactions shown in (a) lanes 2 and 3. (c) CDC34 charging assays were performed as described in the Materials and methods in the presence of RAG1[264–389] or RAG1 storage buffer as indicated. The percentage CDC34 charged was determined by western blot. The average and standard deviation for three independent trials are shown. R1, RAG1.