Western Blot for the High School Classroom

Developed by: Melissa Bunch and Jim Woodbury
© American Association of Immunologists 2001

Overview
This protocol was written to be utilized in a new molecular biology research class for high school students. If can also very well fit into an AP Biology course when studying proteins or the immunological responses and into a biotechnology course.

This protocol follows a complete study of DNA, RNA and proteins. At this point the student has already become familiar with such techniques as using a digital micropipette, metric conversions using units such as microliters, calculating molarity and making solutions.  This exercise will allow the student to:

• Transfer the protein from a polyacrylamide gel to a nitrocellulose membrane
• Understand the antigen/antibody complex
• Develop the membrane by using a color-change technique
• Analyze the membrane to verify presence of protein
• USe a TI-83 calculator to estimate the size of the protein (band)