Protein analysis: Western Blotting

Imagine that you are only interested in one specific protein, and trying to detect its presence across hundreds of samples. Each sample is from a different batch of cells, and there are bound to be other proteins or mixture of contaminants that will limit your ability to detect the protein of interest. The Western Blotting technique is a sophisticated method of protein analysis, as compared to a more global detection method like Coomassie staining. Both techniques can be used to estimate protein size, when coupled with SDS-PAGE, but vary in their specificity and sensitivity.

#biolabcollective #protein #biochemistry #microbiology #laboratory #teaching

Timestamps:
0:00 Introduction
0:35 Western Blotting
2:15 Membrane Transfer
2:47 Antibody Binding
3:36 Making sense of the results
4:54 Conclusion

Presenter: Jack Wang

[jayzeeeasy]

Wouldn’t the tertiary structure of a target enzyme like LDH be broken down during SDS into strands of amino acids bound to SDS therefore disallowing the relevant immunological markers to bind? I assume the markers can only bind to enzymes in their natural structure, is this so?