Immunodetection

Western blot analysis

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Explore the principles behind the technique of western blotting and learn how to calculate, quantify and normalise unknown proteins using a Bradford assay. Learn to identify common western blot errors using stylised reproductions and then identify those same errors with real blot images. Interpret western blot data in novel research-based scenarios.

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Learning outcomes

Recall the principles behind the methodology and analysis of western blotting.
Calculate the appropriate dilutions for a Bradford assay.
Construct and use a standard curve to calculate protein concentration and appropriate volumes.
Evaluate western blot data to deduce common errors and propose experimental conclusions.

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Skills

Designing and Planning Experiments

Determining the reasons for not using a freezer stock to make immediate working solutions

Manipulating and Presenting Data

How to calculate working dilutions and subsequent concentrations needed for a Bradford assay
How to build a standard curve by defining the axis titles and ranges
How to use the equation of a standard curve to interpret novel absorbances from protein samples
How to calculate specific volumes needed to reach a normalised protein concentration for a western blot experiment

Interpreting and Analysing Data

Interpreting a Bradford assay and determining whether the protein extraction was useable in a western blot experiment
Identifying different western blot errors from stylised and real western blot images
Interpreting western blot results from novel case studies

Concepts, Principles and Theories

Defining the purpose of western blot as an analytical technique and what it measures
Distinguishing the different methods of western blot and the procedures and components associated therewith

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Details

Degrees

Biomedical science

Biological science

Pharmaceutical science

Modules

Molecular biology

Topic

Immunodetection

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