A growing team of groundbreaking scientists around the world are now sharing their lab notebooks online
It’s being a while since I wrote my first post. I have been busy in the lab trying to isolate monomeric and stable samples of the N-HEAT domain just as we demonstrated with the C-HEAT domain (and by Dr. Rachel Harding). In this post, I summarize the results of the first round of purification of Read More …
In my previous post I mentioned that there is controversy if HDAC11 can deacetylate histones. I decided to look at the effect of overexpression of HDAC11 on exogenous histone acetylation in cells. In my preliminary results I noticed that exogenous GFP-tagged histones, one day post transfection, have very low levels of acetylation; therefore I decided Read More …
Hi all, today’s post covers some concepts that I have previously described: expression and purification of protein constructs, and testing ligands in a displacement assay. If you’ll recall, I tried to develop and optimize a fluorescence polarization (FP) displacement assay for the ZnF-UBD of USP5 for some time but found that FP was a no Read More …
In my previous post https://openlabnotebooks.org/protein-protein-docking-for-protac-discovery/ I showed that HADDOCK was the best protein-protein docking tool among those I tested to predict how E3 ligases interact with their protein substrates. Here, I ask whether docking virtual libraries of PROTAC candidates to these E3 ligase – substrate protein interfaces can be used to predict which PROTACs are active. Read More …
Purifications of HTT and HTT-HAP40 from insect (Sf9) and mammalian (EXPI293F) expression systems
Summer travels showcasing the huntingtin protein toolkit
Back again. Two posts in one month! I know its crazy, but I’m just excited to tell you about the progress that is being made on this project. First things first, purifications. This week I have four more to tell you about. The first is GFP, and unlike all of my other posts, this is Read More …
Hello again! It has been a while since my last post but that is not for lack of data. Over the past month I have been purifying like crazy, adding more proteins to out bank of possible interaction partners. As we are now in the final month of the project, I want to take this Read More …
My main project revolves around the E3 ligase UBE3A (I will be writing a project overview for this later), but I am also continuing some work in the same vein as Jolene Ho’s work looking at role of HTT in neurological disorders. HTT encodes a protein called huntingtin whose function is not known but appears Read More …
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 …
