Purifications of HTT and HTT-HAP40 from insect (Sf9) and mammalian (EXPI293F) expression systems

I am really excited to be working in collaboration with so many great groups in the Huntington’s disease research space. One of my contributions to many of these collaborations is to provide huntingtin (HTT) and HTT-HAP40 protein samples with different polyglutamine tract lengths. My collaborators can then use the material to investigate both the structure Read More …

Successful generation of fragments of the HTT protein and improving the purification procedure for the HTT-HAP40 complex

It has now been almost 2 years since I set out to try and make fragments of the huntingtin protein which might be amenable to structural analysis with X-ray crystallography. X-ray crystallography is a fantastic method and allows us to see the molecules in very fine atomic detail which is important if we are to Read More …

Optimisation of HTT-HAP40 purification using heparin affinity chromatography

In order to do any biochemical studies of the huntingtin protein, I must first purify the protein in large amounts. The current protocol for protein purification is adapted from other published protocols and requires a long incubation of clarified cell lysate with FLAG resin which means my protein is sitting around with proteases and other Read More …

Manuscript finished! Design and characterisation of mutant and wild-type huntingtin proteins produced from a toolkit of scalable eukaryotic expression systems

Hi folks! This past month or so I have been busy getting a manuscript draft prepared. This article details the HTT constructs we have made and shared with the community, how they can be used to make pure huntingtin protein of different polyQ lengths and characterisation of the protein samples that we made. We think this Read More …