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. Author manuscript; available in PMC: 2009 Dec 1.
Published in final edited form as: J Am Soc Mass Spectrom. 2008 Aug 12;19(12):1875–1886. doi: 10.1016/j.jasms.2008.08.004

Figure 2. Preparation of the crude NADH dehydrogenase (NDH, Fp subcomplex) from carbamoylmethylated complex I.

Figure 2

Complex I was alkylated with iodoacetamide according to the procedure described in Experimental Methods. 100 % ethanol was added to a final concentration of 9% (v/v), and the mixture was incubated at 40 °C for 10 min. The sample was then chilled in an ice-salt water bath for 10 min and subjected to centrifugation at 25,000 rpm for 30 min. The supernatant containing crude NDH was collected and concentrated with Centricon 30. Carbamoylmethylated complex I and crude NDH were subjected to SDS-PAGE using 4–12% Bis-Tris polyacrylamide gel. Lane 1, Carbamoylmethylated complex I (30 μg); lane 2, crude NDH preparation (15 μg) from carbamoylmethylated complex I. M represents a molecular weight marker.