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. Author manuscript; available in PMC: 2015 May 4.
Published in final edited form as: Biochem J. 2014 Dec 1;464(2):251–258. doi: 10.1042/BJ20140845

Figure 3. Associations amongst Munc18-1, Doc2b and Munc18c in GST interaction assays.

Figure 3

Bacterially-expressed GST-tagged-Munc18-1 and -Munc18c proteins were purified and linked to sepharose beads. GST-Munc18-1 (A) or GST-Munc18c (B) were used in interaction assays using reactions containing equimolar amounts of soluble untagged recombinant proteins in the presence or absence of Doc2b, and were compared against binding to the GST protein alone. Coprecipitated proteins were resolved by 10% SDS-PAGE for immunoblot detection (IB). Ponceau S staining was used to gauge equivalent loading of GST-tagged proteins. Input proteins were sampled from reactions to confirm the presence of each protein in the reaction. Data are representative of at least three independent experiments using two or more independent batches of recombinant proteins.