Reduction of HTT Protein through Chemical Inhibition

The experimental set-up and results covered in this post can be found here: 10.5281/zenodo.1495129. Apologies for the long delay between this post and my last post, it really has been quite some time! As postdocs, we generally have several projects running simultaneously and we have to split our time between these projects. I am no Read More …

Generating HTT Knockdown Models

The experimental set-up and results covered in this post can be found here: doi.org/10.5281/zenodo.1420537 I was previously using an HTT overexpression model to look at any changes in proteins related to HTT or its function. Unfortunately, none of the related proteins I looked at showed any significant change in protein levels with HTT overexpression. There Read More …

Checking Disruption of Proteins of Interest with Overexpression of HTT

The experimental set-up and results covered in this post can be found here: 10.5281/zenodo.1323902 As I’ve been having problems overexpression HTT properly in my cancer cell lines, I decided to continue looking at other proteins of interest using our best overexpression model so far, which is in HEK293T cells. Although it may not be a cancer Read More …

Overexpression of Expanded HTT in Brain Cancer Cell Lines

The experimental set-up and results covered in this post can be found here: 10.5281/zenodo.1292884. A few posts back, I tried expressing HTT with different lengths of Q repeats in mammalian cells using baculovirus (read more here). However, expression was not very good, especially for HTT with longer Q repeats (have a look at these results here). Read More …

Visualisation of Purified HTT Proteins (2)

The experimental set-up and results covered in this post can be found here: 10.5281/zenodo.1253365. Recently, I helped run some Western blots for Rachel Harding, a fellow postdoc and open notebooker at SGC (read more about her work here: http://labscribbles.com/), for a publication on a toolkit to enable people to more easily purify and study HTT (you Read More …

Visualisation of Purified HTT Proteins

The experimental set-up and results covered in this post can be found here: 10.5281/zenodo.1233602.   Lately, I’ve been helping Rachel Harding, a fellow postdoc at SGC and one of the pioneers of the Extreme Open Science Initiative at SGC, with some HTT experiments. She is also working on HTT, and has in fact been working on Read More …

N-terminal vs C-terminal FLAG-tagged HTT

The experimental set-up and results covered in this post can be found here: 10.5281/zenodo.1202292 In my previous post, I used baculovirus to overexpress HTT in mammalian cells. Although I managed to overexpress HTT, this HTT is actually FLAG-tagged, and very strangely, I could not detect FLAG! Let me first explain what a FLAG-tag is. FLAG is Read More …

Overexpressing HTT in mammalian cells

The experimental set-up and results covered in this post can be found here: 10.5281/zenodo.1172644   One of the main aims of my project is to understand the role of normal and mutant HTT in mammalian cells. To do so, I will be overexpressing HTT in mammalian cells using baculovirus. The use of baculovirus for protein expression Read More …