Large scale purification of full-length huntingtin Q23, Q46 and Q78 from insect cells

Hello lab scribbles readers! I have been busy in the lab making lots of protein for some upcoming electron microscopy experiments – all materials and methods can be found on Zenodo. These proteins are the complete full-length human huntingtin protein with either 23, 46 or 78 glutamine amino acids in the polyQ repeat stretch representing Read More …

Test Expression of Huntingtin Domain Constructs in Insect Cell Culture

I am back from vacation and other distractions and now have some new and exciting data to share with everyone. Following on the previous limited proteolysis experiment where I chopped up the protein into proteolyticaly stable nuggets which allowed me to work out the potential domain boundaries of the huntingtin protein, I designed constructs which Read More …

Limited proteolysis of full-length huntingtin with chymotrypsin

I have received data from SPARC, Sick Kids Hospital Toronto, for the limited digest experiment I previously wrote about. This experiment aims to work out where the flexible linker regions of the protein structure are, therefore predicting which regions of the protein sequence correspond to folded domains.All of the experimental data is uploaded onto Zenodo. Very Read More …