An active fraction was subjected to 15 % (w/v) polyacrylamide-gel electrophoresis as described by Bollage and Edelstein (1991 ) using a Tris–Tricine buffer system performed according to the manufacturer’s instruction of the Mi-Protean® Tetra system (Bio-Rad, USA) at 100 V for 45 min. Fermentas Spectra™ Multicolor Low Range Protein Ladder (Thermo Scientific, USA) was used to estimate the molecular weight of the pure bacteriocin. The gel was stained using a Pierce® Silver stain kit (Thermo Scientific, USA).
Fermentas spectra multicolor low range protein ladder
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Fermentas Spectra™ Multicolor Low Range Protein Ladder is a pre-stained protein marker used for monitoring protein separation during SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). The ladder contains a mixture of proteins with known molecular weights, allowing for the estimation of the molecular weights of unknown protein samples.
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2 protocols using fermentas spectra multicolor low range protein ladder
1
Bacteriocin Molecular Weight Determination
2
Protein Separation by SDS-PAGE
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Polyacrylamide gel electrophoresis was performed to separate proteins. Briefly, 10 μg protein per well was loaded in 15% (w/v) polyacrylamide-gels and separation was achieved at 100 V for 45 min using a Tris/glycine/sodium dodecyl sulfate buffer (Mini-Protean Tetra system, Bio-Rad, Hercules, CA). The molecular weights of proteins were estimated using the Fermentas Spectra Multicolor Low Range Protein Ladder (Thermo Fisher Scientific, Waltham, MA). The gel was stained using Bio-Safe Coomassie G-250 (Bio-Rad).
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