Formate hydrogenlyase in the hyperthermophilic archaeon, -2

The first (S2) and second (S3) wash of membrane fraction as well as the washed membrane fraction (WM). Proteins were separated on 12% SDS polyacrylamide gel after boiling in SDS-loading buffer for 10 min and the FdhB subunit was monitored using anti-FdhB antibody. One thousandth of the volume of the prepared fractions was loaded onto the gel to make the signals comparable. . Proteins were separated on 6% native gel and stained for hydrogenase activity. S: soluble fraction M: unwashed membrane fraction WM: washed membrane fraction. The buffer used for the washing step contained 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. . The proteins from the activity stained gel were transferred to nitrocellulose membrane and screened with anti-FdhB antibody. The hydrogenase activity band corresponding to the FdhB signal is indicated with an arrow.Copyright information:Taken from "Formate hydrogenlyase in the hyperthermophilic archaeon, "http://www.biomedcentral.com/1471-2180/8/88BMC Microbiology 2008;8():88-88.Published online 3 Jun 2008PMCID:PMC2432063.