
Updates on all the huntingtin samples I have made and what has been keeping me busy in the lab
Hi folks! It has been quite a while since I last updated you all on what I have been up to in the lab. It has been a busy time these past few months and I have not found as much time as I would like to keep LabScribbles up to date. Better late than Read More …
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 …
Summer travels showcasing the huntingtin protein toolkit
I have been really lucky this summer to have spent a lot of time travelling and attending lots of different meetings. This has been a fantastic opportunity for me to spread the word about our publication earlier this year in the Journal of Biological Chemistry which outlines the toolkit we made to allow other labs Read More …
Identifying new members of the huntingtin interactome with BioID and other updates
I have been busy in the lab working at the bench and training our new recruits to the Huntington’s disease research team! Welcome Jacob and Claudia! Using a technique called BioID, we have identified proteins which are proximal or nearby to the HTT protein in cells. To date, no published literature details the use of 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 …
Update on meetings, manuscripts and mighty cool experiments…
It has been a while since I have written an update on here and there is a lot to update you all on. What has taken up a lot of my time has been work on our huntingtin toolkit manuscript which was recently accepted at the Journal of Biological Chemistry – hoorah! I am still 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 …
Huntington Society of Canada 2018 Conference in Kelowna
Hi everyone! This weekend I am in rainy Kelowna for the Huntington Society of Canada National Conference. I am excited to meet all the participants and hear from scientists, clinicians, patients and other folks who attend this important meeting. On Saturday, I will be giving a short presentation on free and legal ways to access Read More …
Biophysical investigation of HTT and HTT-HAP40 using DSLS and SAXS
Hi folks! Over the past month or so, I have been conducting biophysical characterisation experiments of the HTT and HTT-HAP40 samples I am able to purify in the lab. I have used DSLS which measures thermal stability by monitoring protein aggregation over a temperature gradient. In an ongoing collaborative effort with Sasha Lemak and Lixin Read More …