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. Author manuscript; available in PMC: 2010 Jul 15.
Published in final edited form as: Anal Biochem. 2009 May 3;390(2):221–223. doi: 10.1016/j.ab.2009.04.031

Figure 2.

Figure 2

(A) Schematic of DTDP-protection of S105 Cys51 sulfhydryls. DTDP was included in the membrane extraction buffer prior to purification, and the thiol-specific reagent was removed from the purified protein with the reducing agent, TCEP. Cys51 was fully accessible to PEG-mal once the DTDP had been removed. (B) Western blots of PEGylation samples from S105 and S68 purifications, detected with antibodies directed against peptides corresponding to the C-termini of S105 [7] or S68 [9]. Lanes 1-4, DTDP-protection of S105 thiols during membrane extraction and protein purification. Lanes 5 and 6, purified S2168G48Chis without DTDP included in the membrane extraction buffer; lanes 7-10, purified S2168G48Chis with DTDP included in the membrane extraction buffer (* indicate the monomer bands, ** indicate the dimer bands, and → indicate the PEGylation products).