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. Author manuscript; available in PMC: 2016 Mar 1.
Published in final edited form as: Plant J. 2015 Mar;81(5):723–735. doi: 10.1111/tpj.12762

Figure 7. Preparative separation and immunoblot analysis of Lb derivatives generated by nitration.

Figure 7

(a) Separation of Lb derivatives on preparative native gels. PvLba (upper gels) and GmLba (lower gels), both at 1 mM, were treated for 30 min with 10 mM H2O2 + 10 or 100 mM NO2− in 20 mM potassium phosphate buffer (pH 6.5). The protein products (bean, B1 to B7; soybean, S1 to S7) were eluted from gels and analyzed by MS.

(b) Immunoblot analyses of proteins eluted from (a) using antibodies against NO2-Tyr and Lb. Besides monomeric nitrated Lb, dimers and other Lb aggregates (>27 kDa) were formed with different levels of nitration. GmLba was more resilient to nitration than PvLba, as the former showed less intense signal with the NO2-Tyr antibody and maintained most Lb in monomeric form.