Learning Outcomes:
On completion of this module, students are expected to be able to:
1. Describe the basic principles of cellular protein production and protein quantification with an emphasis on Western Blotting. These include, but are not limited to, the following topics:
a. Gene transcription and translation
b. Intracellular protein processing and post-translational modifications
c. Protein isolation
i. Cell cultures vs tissue
ii. Protein stability
d. Quantification of total protein
i. Bradford assay, BCA assay
e. Protein separation techniques
i. Polyacrylamide gel electrophoresis
1. Buffer selection and gel density
2. Denaturing or native conditions
f. Transfer of separated proteins to membrane
g. Protein staining techniques
i. In gel staining (coomassie)
ii. Non-specific protein stain (Ponceau)
iii. Antibody-mediated staining
h. Protein quantification
i. Importance of normalising for protein loading
ii. Limitations of dynamic range
2. Demonstrate laboratory competence in performing tasks relating to Learning Outcome 1.