Proteins were fractionated by performing SDS-PAGE (4–20%) stained with Coomassie blue (Euromedex, Souffelweyrshim, France). For immunoblot analysis, the proteins were transferred to nitrocellulose membranes before being revealed with anti-Histidine monoclonal antibodies (Qiagen). Immune complexes were detected with anti-rabbit peroxidase-conjugated secondary antibodies (Promega) and enhanced chemiluminescence reagents (Pierce, Illkich, France).
Coomassie blue
Manufactured by Euromedex
Sourced in France
Coomassie blue is a synthetic dye used in various laboratory techniques, primarily for the staining and detection of proteins in polyacrylamide gels. It is a versatile and widely used reagent in biochemistry and molecular biology.
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2 protocols using coomassie blue
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SDS-PAGE Protein Fractionation and Immunoblotting
2
SDS-PAGE and Immunoblot Analysis
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Proteins were fractionated by performing SDS-PAGE (12% except where indicated) stained with Coomassie blue (Euromedex, Souffelweyrshim, France). For electrophoresis under non-reducing conditions, β-mercaptoethanol was omitted from the SDS-PAE gels, and the proteins were not heated before loading into the gel. For immunoblot analysis, the proteins were transferred to nitrocellulose membranes before being revealed with specific polyclonal antibodies. Immune complexes were detected with anti-rabbit peroxidase-conjugated secondary antibodies (Promega) and enhanced chemiluminescence reagents (Pierce, Illkich, France). Anti-HetR antibodies were developed by Covalab and used at a 1:1000 dilution.
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