Nupage mes sds

SDS-PAGE analysis of the protein hydrolysates was performed on NuPAGE® Novex 4-12% Bis-Tris Gels using the XCell-sure lock Mini-Cell (Invitrogen, Madrid, Spain). Electrophoresis was carried out at 200 V, and the running and sample buffers used were NuPAGE® MES-SDS, and NuPAGE® LDS (Invitrogen), respectively. Runs were carried out under non-reducing conditions in which 2-mercaptoethanol was omitted in the denaturing buffer. Electrophoretic bands were stained with SimplyBlueSafeStain (Invitrogen), followed by destaining in deionized water. The molecular weight of poly-and oligopeptides was determined by comparison with the molecular weight marker solution Mark 12 TM (Invitrogen) containing myosin (200 kDa), -galactosidase (116.3 kDa), phosphorylase B (97.4 kDa), bovine serum albumin (66.3 kDa), glutamic dehydrogenase (55.4 kDa), lactate dehydrogenase (36.5 kDa), carbonic anhydrase (31.0 kDa), trypsin inhibitor (21.5 kDa), lysozyme (14.4 kDa), aprotinin (6.0 kDa), insulin B chain (3.5 KDa) and insulin A chain (2.5 kDa).

Protein profiles of lentil soluble fractions were analyzed by SDS-PAGE. Electrophoresis was carried out loading 20 µg of protein/well on NuPAGE ® Novex 4-12% Bis-Tris Gels (Invitrogen, Madrid, Spain). Gels were placed in XCell-sure lock Mini-Cell (Invitrogen, Madrid, Spain) and run under reducing conditions at 200 V. NuPAGE ® MES-SDS and NuPAGE ® LDS (Invitrogen, Madrid, Spain) were used as running and sample buffers, respectively. Gels were stained with SimplyBlue SafeStain (Invitrogen, Madrid, Spain) for 1 h and distained in deionized water for 2 h. The molecular weight of poly-and oligopeptides was determined by comparison with the molecular weight marker Novex® Sharp Prestained Protein Standard (Invitrogen, Madrid, Spain) containing proteins from 3.5 to 260 kDa.