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. Author manuscript; available in PMC: 2008 Aug 15.
Published in final edited form as: Chem Res Toxicol. 2006 Oct;19(10):1366–1373. doi: 10.1021/tx060106t

Fig. 4.

Fig. 4

Immunoblot of pooled human liver microsomes, probed for both NADH cytochrome b5 reductase (A) and cytochrome b5 (B) content. Lane 1: pooled human liver microsomes, lane 2: duplicate pooled human liver microsomes, lanes 3−5: increasing concentrations of purified recombinant b5R (Panel A; 0.18, 0.45, and 0.72 pmol) or cyt b5 (Panel B; 1.2, 2.94, and 8.82 pmol) to establish standard curves (pmol b5R = 0.0166 * HLM b5R density + 0.2198; pmol cyt b5 = 0.6455 * HLM cyt b5 density + 0.6409). (Note: this cyt b5 antibody has a lower affinity for cyt b5 (less dense bands), compared to the b5R antibody for b5R; therefore, more pmol of cyt b5 have been loaded as standards to give a linear range for quantitation). Results from four immunoblots (representative shown) indicated a native b5R : cyt b5 molar ratio of between 1:8 to 1:10 in HLM.