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. Author manuscript; available in PMC: 2014 Jun 2.
Published in final edited form as: Mol Microbiol. 2011 Jul 29;81(5):1390–1401. doi: 10.1111/j.1365-2958.2011.07770.x

Figure 4.

Figure 4

Incubation of recombinant LF with GRP78 at pH 7.5 results in its conversion from a trypsin resistant to sensitive conformation in vitro. LF (100 ng; List Biologicals) was incubated in the presence or absence of GRP78 (538 ng; Enzo Life Sciences) in unfolding buffer (25 mM Tris-HCl, 150 mM NaCl, 10 mM KCl, pH 7.5) for 10 min at 37°C, before the addition of trypsin (2.5 ng; Promega) or buffer alone and then incubated for 1 hr at 37°C. Digestion reactions were stopped by addition of 2X SDS-PAGE loading buffer with β-mercaptoethanol and boiled for 5 min prior to electrophoresis and detection on immuno-blots probed with anti-LF (Pierce Antibodies). Pixel density was assessed using Kodak 1D v.3.6.0 software. *Statistical analysis was performed using GraphPad Prism v.5.01. ANOVA followed by the Tukey Test revealed p = 0.0002 (n = 7).