A growing team of groundbreaking scientists around the world are now sharing their lab notebooks online
Hello again! In previous posts I talked about how we were using a co-culture system to grow our leukemic cells. Although this is a great way to maintain cells, it can complicate some analyses where we cannot distinguish mouse (stroma) from human (AML) cells. We wanted to determine H3K27me3 levels (the mark that EZH2 is Read More …
In nanoBRET assay, a fluorescently labelled tracer binds to the ATP-binding pocket of ACVR1/ALK2. This binding brings the tracer into the proximity (<10nm) of Nano-Luciferase which is fused to the C-terminus of ACVR1/ALK2. Bioluminescence resonance energy transfer (BRET) occurs where the energy released by Nano-Luciferase is transferred to the fluorophore via intermolecular forces. Nano-Luciferase and Read More …
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 …
After the disappointment of looking through ~13000 crystal drop images over the past couple of weeks and finding pretty much nothing in most of them and nothing that actually diffracted nicely in the rest, it was incredibly heartwarming and hugely inspiring to attend the FOP family and friends conference in Manchester last weekend. FOP Friends is Read More …
A major therapeutic avenue for chordoma is targeting the receptor tyrosine kinase, EGFR. Its ligand, EGF, is overexpressed as a transcriptional target of Brachyury and polysomy of EGFR’s region of chromosome 7 is frequent in chordoma. To this extent I, in collaboration with SGC-UNC’s post-doc Chris Asquith, am working on the profiling of a number Read More …
Deposition: In the past two weeks my work has mostly been full of dealing with structures. The first thing I needed to do was deposit some structures of ALK2 co-crystallised with three drug molecules that I had solved for our collaborators Swen Hoelder and Liam Hudson at ICR, who had been developing compounds against ALK2. Read More …
Another small set of compounds was recently synthesized, the major change in Set2 is the substitution of the sulfur atom by oxygen or nitrogen. The change was made in position 6 of the pyrimidine ring. These compounds along with others from Set1 are currently under screening in the NanoBRET cellular assay (performed by Carrow Wells).
Full-length huntingtin protein aggregation and first attempts to co-purify HTT-HAP40 from insect cells
Greetings! My first post here will be a brief introduction. More in-depth experiment and data posts are coming soon! I am a post-doctoral research associate in the UNC Catalyst for Rare Diseases1. Throughout my education and training, I have been interested in cancer – I worked on epigenetics of ovarian cancer as an undergraduate and Read More …
I have had little success with DSF and FP assay development for USP5 Zf-UBD to identify small molecule ligands of this domain. I decided to try 19F NMR as a potential screening assay. NMR spectroscopy can be used to visualize changes in protein spectra upon addition of low complexity molecules as well as to quantify Read More …
