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Figure 4 | BMC Microbiology

Figure 4

From: Formate hydrogenlyase in the hyperthermophilic archaeon, Thermococcus litoralis

Figure 4

Activity staining (A) and immunoblot analysis (B) of the CHT chromatography fractions having hydrogenase activity. A. Proteins were separated by 5–13% gradient CN-PAGE and stained for hydrogenase activity. The fractions of the CHT chromatography were concentrated (Millipore Amicon® Ultra centrifugal filter devices, cut-off 100 kDa) then the buffer was replaced by 1% dodecyl-β-D-maltoside and 750 mM 6-aminohexanoic acid in 50 mM Bis-Tris pH 7.0 The unsoluble materials were removed by centrifugation at 50000 × g for 20 min and the supernatants were loaded onto the gel. B. Proteins were separated on 12% SDS polyacrylamide gel after boiling in SDS-loading buffer for 10 min, transferred to nitrocellulose membrane and screened with anti-FdhB antibody. M fr.: membrane fraction that was loaded onto the CHT column. The numbers mean the numbers of the fractions analyzed.

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