PHYS40110 Analysis of protein expression - Western Blot

Academic Year 2023/2024

This module is designed to provide an introduction to antibody-based techniques for quantifying protein expression in samples derived from cultured cells or tissues with an emphasis on assay design and development. The primary aim of this module is to give students hands-on laboratory experience in Western Blotting, and an understanding of the various controls that are necessary to ensure reliable data.

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Curricular information is subject to change

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.

Student Effort Hours: 
Student Effort Type Hours
Tutorial

12
Laboratories

60
Autonomous Student Learning

40
Total

112
Approaches to Teaching and Learning:
This module involves approximately ten weeks of laboratory-based practical activity.
Students will work closely with a research active supervisor.
Teaching will involve laboratory work; active/task-based learning; critical writing; reflective learning; enquiry & problem-based learning.
 
Requirements, Exclusions and Recommendations

Not applicable to this module.


Module Requisites and Incompatibles
Not applicable to this module.
 
Assessment Strategy  
Description Timing Open Book Exam Component Scale Must Pass Component % of Final Grade
Continuous Assessment: Laboratory Report Varies over the Trimester n/a Graded Yes

100

Carry forward of passed components
No
 
Remediation Type Remediation Timing
Repeat Within Two Trimesters
Please see Student Jargon Buster for more information about remediation types and timing. 
Feedback Strategy/Strategies

• Feedback individually to students, post-assessment

How will my Feedback be Delivered?

Individual feedback will be available after the laboratory reports have been graded.

Timetabling information is displayed only for guidance purposes, relates to the current Academic Year only and is subject to change.
 

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