Use of Two Dimensional Semi-denaturing Detergent Agarose Gel Electrophoresis to Confirm Size Heterogeneity of Amyloid or Amyloid-like Fibers

The overall goal of this two-dimensional semi-denaturing detergent agarose gel electrophoresis is to confirm size heterogeneity of amyloid or amyloid-like fibers seen during traditional SDD-AGE. This method can help answer key questions in the amyloid or protein aggregation fields, such as whether size heterogeneity seen during traditional SDD-AGE is the native state of the fibers in vivo or the result of protein degradation or dissociation during gel electrophoresis. The main advantage of this technique is it can be performed easily in lab without specialized equipment.

No additional equipment is required beyond what's used in traditional SDD-AGE. First, seed the amyloid-producing HT-29 colon cancer cells in a 10-centimeter tissue culture dish. Then, incubate the cells overnight at 37 degrees Celsius with 5%carbon dioxide.