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. Author manuscript; available in PMC: 2013 Mar 1.
Published in final edited form as: Protein Expr Purif. 2011 Dec 8;82(1):106–115. doi: 10.1016/j.pep.2011.11.016

Figure 2. Comparison of the biotinylation efficiency of BirA in different cellular compartments of S2 cells.

Figure 2

Conditioned media from the cells, expressing Avi-GCPII and BirA (targeted to three different cellular compartments - cytoplasmic, secreted, or within the ER), were harvested 3 days post-induction and analyzed on Western blot. Western blots were visualized with the monoclonal antibody GCP-04, which recognizes total Avi-GCPII (Panel A), and Neu-HRP, which recognizes only the biotinylated Avi-GCPII (Panel B). 1. 7 ng (Panel A) or 3 ng (Panel B) of Avi-GCPII standard; 2. medium from cells with BirA localized within the ER (stable cell line 1); 3. medium from cells with BirA localized within the ER (stable cell line 2); 4. medium from cells with secreted BirA; 5. medium from cells with BirA localized in cytoplasm. Lanes 2–5 in both panels contained identical volumes of the analyzed samples.