Journal: Microorganisms

Article Title: Molecular Cloning and Characterization of a New Family VI Esterase from an Activated Sludge Metagenome

doi: 10.3390/microorganisms10122403

Figure Lengend Snippet: ( A ) SDS-PAGE analysis of overexpressed and Profinity eXact purified Est6 in E. coli BL21 Star (DE3). Samples were run on a 4–12% NuPAGE Novex Bis-Tris mini gel. Lane M, protein molecular mass ladder (Fermentas); lane 1, total soluble cell protein; lane 2, Est6 in 50 mM sodium phosphate buffer (pH 7.0); lane 3, Est6 in 50 mM Tris-HCl buffer (pH 8.0). The arrows indicate the Profinity eXact tagged Est6 and tag-free Est6, respectively. ( B ) Native PAGE analysis of purified Est6. Samples were run on a NativePAGE Novex 3–12% Bis-Tris gel. Lane M, native protein mass ladder (NativeMark unstained protein standard, Invitrogen). Lane 1, Est6 in 50 mM sodium phosphate buffer (pH 7.0); lane 2, Est6 in 50 mM Tris-HCl buffer (pH 8.0). ( C ) Zymogram of purified Est6 on an agarose gel containing 1% tributyrin. The sample order is as same as that in ( B ).

Article Snippet: The recombinant protein was purified with a Profinity eXact purification resin (Bio-Rad) in accordance with the manufacturer’s protocol.

Techniques: SDS Page, Purification, Clear Native PAGE, Agarose Gel Electrophoresis