Previously I reported some exciting test expression data where we showed that small chunks of the huntingtin protein could be made in the lab, a key milestone to aid future experiments where we try and work out what the huntingtin protein looks like and what it might be doing in the cells of our bodies. Since then, I have been completing some further work to tie up some loose ends and validate the previous data.
This has included:
- rerunning some of the SDS-PAGE to more clearly see the bands of protein of interest
- completing some mass spectrometry experiments to determine if the blue protein bands do correspond to the huntingtin protein or whether they are an artifact of the experiment
- scaling up the expression of the positive hits to make more of the protein in the coming weeks
Below is an example of some of the mass spectrometry data reported where we showed that the protein bands we see on the gel do correspond to the huntingtin protein; peptides detected are aligned with the huntingtin sequence. This work was completed by Jonathan Krieger, SPARC, Sick Kids Toronto. A big thanks to him for his help!
A detailed write up including all of the materials and methods, all of the raw data and my analysis can be found on Zenodo, open access and available to all. Feel free to get in touch if you have any questions or comments.