Optimizing an Immunohistochemical Protocol for the Analysis of Streptozotocin Effects on Insulin Signaling in Rat Brains

Avery M. Schroeder
Dr. Daniela Ostrowski, Faculty Mentor

Immunohistochemistry is a staining process that allows for the visualization of cellular proteins. My study optimized an immunohistochemical protocol that will be used to analyze the effect of Streptozotocin (STZ), an Alzheimer’s phenotype-inducing drug, on the insulin signaling pathway in the brains of rats. Two primary experimental variables were evaluated. First, the utilization of an antigen retrieval method (heating tissue in a 95 degree Celsius water bath for 20 minutes) was tested. Heat treatment is used to improve antibody binding for proteins that may have been modified during the fixation process. In addition, two buffer conditions were tested (Tris buffer, pH 9 versus Citrate buffer, pH 6). This is relevant, as antibodies have different binding properties at different pH values. My study concluded that tissue heating in combination with the use of a Citrate buffer gives rise to the best staining results. 

Keywords: Immunohistochemistry, Alzheimer's Disease, Streptozotocin, Insulin Signaling

Topic(s):Biology

Presentation Type: Poster

Session: TBA
Location: TBA
Time: TBA