A solution of fibrils was mixed at 3:1 ratio with 4X lithium dodecyl sulfate sample buffer (Thermo Fisher Scientific) and heated at 95 °C for 10 min. Proteins were separated on a NuPAGE 4–12 % Bis-Tris gradient gel (Thermo Fisher Scientific) using NuPAGE MES LDS running buffer (Thermo Fisher Scientific). The gel was stained for 1 h with a solution of 2.5 g/L Coomassie brilliant blue R250 in 20% (v/v) ethanol and 10% (v/v) acetic acid. The gel was destained in 30% (v/v) ethanol and 10% (v/v) acetic acid.
Nupage mes lds running buffer
Manufactured by Thermo Fisher Scientific
NuPAGE MES LDS running buffer is a laboratory reagent used in the electrophoresis of proteins. It is designed for use with NuPAGE Bis-Tris precast gels and the NuPAGE electrophoresis system.
Automatically generated - may contain errors
3 protocols using nupage mes lds running buffer
1
SDS-PAGE Analysis of Protein Fibrils
2
SDS-PAGE Protein Separation and Visualization
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Proteins were separated on NuPAGE® 4–12% Bis-Tris gradient gels (Thermo Fisher Scientific) using NuPAGE® MES LDS running buffer (Thermo Fisher Scientific). Samples were mixed with 4 x NuPAGE® LDS sample buffer (Thermo Fisher Scientific) and denatured by heating for 10 min at 95 °C. The gels were stained with 2.5 g/l Coomassie brilliant blue R250, 20% (v/v) ethanol and 10% (v/v) acetic acid for 1 h, before they were destained in 30% (v/v) ethanol and 10% (v/v) acetic acid.
3
Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis
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An aliquot of the fibril solution was mixed at a ratio of 3:1 with 4X lithium dodecyl sulphate sample buffer (Thermo Fisher Scientific Co., Waltham, MA) and incubated at 95 C for 10 min. The samples were analysed using a NuPAGE 4-12% Bis-Tris gradient gel (Thermo Fisher Scientific Co., Waltham, MA) and NuPAGE MES LDS running buffer (Thermo Fisher Scientific Co., Waltham, MA). The gel was stained for 1 h in a solution of 2.5 g/L Coomassie brilliant blue R250 containing 20% ethanol and 10% acetic acid. The gel was finally destained in 30% ethanol and 10% acetic acid.
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