Factors Influencing Protein Separation

SDS-PAGE separates proteins primarily based on their size. As proteins can have a wide range of molecular weights, researchers frequently will wish to observe specific proteins more clearly by enhancing the separation of the proteins within a particular molecular weight range. This increased resolution can be achieved by use of different percentages of acrylamide when casting a gel. Additionally, different buffers used to run the gel can affect the protein mobility. Figures 2 and 3 show the migration of proteins in NuPAGE® and Bolt™ gels using different buffers.

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Figure 1. Migration patterns of protein standards on NuPAGE® Novex® gels. 1


* Migration patterns of Novex® Sharp Protein Standards (Cat. No. LC5800, Prestained; Cat. No. LC5801, Unstained) on NuPAGE® Bis-Tris gels.
† Migration patterns of HiMark™ Unstained Standard (Cat. No. LC5688) on NuPAGE® Tris-acetate gels.

Figure 2. Migration patterns of protein standards on Bolt™ Bis-Tris Plus gels.