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. 2005 Apr;14(4):902–913. doi: 10.1110/ps.041048805

Figure 3.

Figure 3.

Formation of MER-45-kDa hGH by reassociation of 22-kDa hGH subunits and prevention of their reassociation by modification with iodoacetamide. An analytical 13.5% SDS polyacrylamide gel in the absence of reductants was used to separate samples and to assess the degree of 22-kDa hGH subunit association. (Section I) MER-45-kDa hGH was dissociated into 22-kDa hGH subunits by incubation in 10% mercaptoethanol at 100°C for 5 h (lane 1). The dissociated 22-kDa hGH samples were then dialyzed against 5 mM ammonium bicarbonate buffer containing 20 mM 2-mercaptoethanol (lane 2) or with buffer in absence of reductant (lane 3). (Section II) MER-45-kDa hGH was dissociated into 22-kDa hGH subunits by incubation in 10% mercaptoethanol at 100°C for 5 h, and 22-kDa hGH subunits were treated with 7 M iodoacetamide for 15 min at room temperature (lane 1). The dissociated and modified 22-kDa hGH samples were then dialyzed against 10 mM Tris-HCl buffer (lane 2).