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Protein Evolution and the Western Blot

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Description

The Western blotting procedure is rapidly replacing conventional methods for identifying and characterizing specific proteins in complex protein mixtures. This technique is used extensively for this purpose in the research laboratory and is increasingly being used in diagnostic medicine for detecting proteins of disease agents such as the structural core proteins of the AIDS virus. Here, students will perform this technique to examine the evolutionary distance between different mammals. Guided by step-by-step instructions, they subject serum proteins from six species to gel electrophoresis and transfer the separated proteins to nitrocellulose membranes. They then use an enzyme-linked immunoassay to compare the extent to which the separated albumins in the serum samples are related to those from cow. This exercise provides an exciting lesson in molecular evolution and introduces your students to one of the most important techniques of molecular biology.

Each of the individual experiments are supplied with the chemicals and laboratory guides needed for 16 students working in pairs.

Materials

If you chose one or more of the experiments from this series – you should also order Electrophoresis Package 2M or Electrophoresis Package 2. Electrophoresis Package 2M provides sufficient agarose, gel stain, and electrophoresis buffers for 1 of the individual experiments in this series (four gels with 15ml of agarose per gel). Electrophoresis Package 2 provides sufficient agarose, gel stain, and electrophoresis buffers for up to 6 of the individual experiments in this series.

If the P variant is ordered, you will need to provide well gels. Precast polyacrylamide gels can be ordered from Bio Rad, (800-424-6723) Their catalog number is 1611177.