Purification of HTT N-HEAT_81-1643 construct with different buffer conditions

In my previous purification of the HTT N-HEAT_81-1643 construct, I showed that an additional heparin step did not remove any traces of nucleic acid material, changed purity or oligomeric state of the sample. Additionally, my DLS results showed that this construct forms large particles in solution. Thus, the DLS results in combination with the gel filtration Read More …

Purification of HTT N-HEAT_81-1643 and initial biophysical characterization

In the second purification round of N-HEAT_81-1643, I added an additional purification step with heparin resin. My aim with doing this additional step was to improve the purity of the HTT N-HEAT_81-1643 sample from nucleic acid material/other proteins. Similar experiments proved successful for full HTT+HAP40 sample (by Dr. Rachel Harding). Our results showed that the Read More …

Purification of the HTT C-HEAT domain

I am very excited to write my first post in open notebook. My post on Zenodo summarizes the results of my first ever purification of a huntingtin (HTT) domain. I was able to purify successfully the HTT C-HEAT domain encompassing residues 2088-3144. Figure 1. Anti-FLAG purification of the HTT C-HEAT domain. Eluted protein is seen Read More …

Project Overview

I joined the group of Dr. Cheryl Arrowsmith 2 months ago. Currently, I am working on the Huntington’s disease project along with Dr. Rachel Harding and Jacob McAuley. During these past two months, I have been learning how to purify huntingtin (HTT) protein and overall getting up to date with the project. A brief introduction Read More …