Figure 2.

Analysis of purified sGC protein. Spectral analysis of the purified human sGC enzyme displayed a typical Soret band (430 nm) and the broad α/β band (540–580 nm) indicated by arrows. (Inset) A total of 250 ng of purified recombinant α^Nβ (lanes 1–3) and α^Cβ (lanes 4–6) sGC was separated by 7.5% SDS/PAGE and analyzed by silver staining (lanes 1 and 5) or by Western blotting. The identity of the upper bands as α-subunit was confirmed by anti-hexahistidine antibody (lanes 2 and 6) and validated by antibodies to α-sGC subunit (lane 3). The identity of the lower band as β-subunit was confirmed by antibodies raised against human β-sGC subunit (lane 4). The different mobility of α^N and α^C subunits reflects the difference in the size of the tag. The sizes of the molecular mass markers are indicated in kDa.