IL-4 Western Blot Analysis of High Fat Diet and Control Diet Sprague Dawley Rats

Abstract

The Lowry Protein Assay is a fundamental process in the ultimate procedure of Western Blotting; it is used to determine the protein content of a solution, which is essential in calculating the necessary concentration of antibodies. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gels were prepared with a 12% resolving gel and 5% stacking gel. After running the SDS-PAGE, we performed a western blot analysis on our samples. Western blot results showed that there was a significant difference between IL-4 expression in our two different animal experimental groups. Animals with a maternal high fat diet showed increased expression of IL-4, but animals with a maternal control diet showed lower expressions of IL-4. Methylated DNA immunoprecipitation also confirmed our results and showed a region of hypermethylation upstream of the IL-4 gene in the control group.