The SDS-PAGE analysis of the subunits occurred at pH 5.0 (low pI band, L) and at pH 5.8 (high pI band, H) in the denaturing IEF.

<p>The gel disk of the low pI band (lane L) was cut out from the IEF gel rod and equilibrated in SDS solution at 60C for 30 min, and that of the high pI band (lane H) at 60C for 15 min before they were loaded on the SDS-PAGE with 13% slab gel in pH 9.2 Tris-HCl. The subunits were showed in blue bands after Coomassie Blue G-250 staining and in fluorescence bands under UV-light at 365 nm after Zn(SO₄)₂ staining. Lane M showed marker proteins and lane PE was the prepared R-PE.