Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2013 Oct 12;3:453–458. doi: 10.1016/j.fob.2013.10.005

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2013 The Authors

This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-No Derivative Works License, which permits non-commercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

PMC Copyright notice

Fig. 2 — In vitro ubiquitination of JosK117-only. All reactions are monitored at t = 0, 3 and 20 h. Lanes 1–3: reaction using commercial enzymes (0.16 μM E1, 8 μM UbcH5a and 1 μM CHIP), 50 μM ubiquitin and 12 μM JosK117-only at room temperature [27]. Lanes 4–6: reaction using commercial enzymes (1 μM E1, 8 μM UbcH5a, 8 μM CHIP), 250 μM ubiquitin and 50 μM JosK117-only. Lanes 7–9: reaction using enzymes prepared in the lab at the final concentrations as in lanes 4–6. Lanes 10–12: reaction using Josephin pre-treated with IAA. Lane 13: purified mono-ubiquitinated JosK117-only. Lane 14: molecular weight marker. Di-ubiquitin and poly-ubiquitin chains are indicated with arrows. The bands of E1, E2 and E3 enzymes are indicated with annotation below each corresponding band. The Addgene clone of UbcH5a used for the production in the lab comprise an initial sequence deriving from cloning which explains the higher MW with respect to the commercial enzyme.