You wish to explore the expression
pattern of the POI protein in various tissues of the centaurembryo.
(POI - Protein of Interest - is a fictional protein, used as an
example in this tutorial.) Your goal is to determine whether
the horse-derived organs and the human
derived organs differ in the expression pattern of POI.
To that end you take a 30
days embryo, dissect the various organs, prepare whole tissue lysates from
the various organs and run samples in two equal polyacrylamide gels, taking
special care to load equal amounts of the lysates in each well. You use
one gel to prepare the blot, and treat it with an anti-POI antibody, and
then with a secondary antibody conjugated to a reporter enzyme.
Immunoblotting
will show you only the POI protein on the gel.
You use the second gel to
perform Coomassie staining. This staining will show ALL proteins on
the gel.
(Detects all proteins) |
(Detects the POI protein) |
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