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. 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. This
exercise requires about two 2-3 hour laboratory sessions. The exercise was
designed for 8 groups of students and includes: sera from cow, horse, sheep,
goat, chicken and donkey, standard proteins, anti-cow albumin (peroxidase
linked), nitrocellulose, blotting paper, protein blot stain, Tris buffer saline
(TBS), TBS + NP40, color development buffer, chloronapthol, hydrogen peroxide,
dishes for blot incubation, and gelatin. Requires four 12 well gels for 8 groups
of students.
Click Here for polyacryamide
electrophoresis packages and gels.
Modern
Biology Inc