Novex nupage precast protein gels

Protein concentration in the supernatants was determined using a Bio-Rad protein assay kit (Bio-Rad, Hercules, CA, USA). Absorbance was measured at 595 nm, and bovine serum albumin was used as the standard. For protein gel electrophoresis, 20 μL of unconcentrated culture supernatant was loaded onto a polyacrylamide gel (Novex NuPAGE Precast Protein Gels, Thermo Fisher Scientific) for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Endoglucanase and endo-1,4-β-xylanase activity in the culture supernatants was determined using an Azo-CM-cellulose assay kit (Megazyme) and an Azoxylan kit (Megazyme), in accordance with the manufacturer’s instructions. All estimates were performed in triplicate assays. The statistical significance of differences between two conditions was analyzed using two-tailed Student’s t test. For all tests, significance was set at a P value of <0.01 (*).

The protein concentration in the supernatants was determined using a Bio-Rad protein assay kit (Bio-Rad, Hercules, CA, USA). Absorbance was measured at 595 nm and bovine serum albumin was used as the standard. For protein gel electrophoresis, 20-μL unconcentrated culture supernatant was loaded onto a polyacrylamide gel (Novex^® NuPAGE^® Pre-cast Protein Gels, Thermo Fisher Scientific) for sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Endoglucanase and endo-1,4-β-xylanase activity in the culture supernatants was determined using an Azo-cm-cellulose assay kit (Megazyme) and an Azo-xylan kit (Megazyme) in accordance with the manufacturer’s instructions. All estimates were performed in three repeated assays. The statistical significance of differences among WT and mutant strains was assessed by one-way analysis of variance.