A growing team of groundbreaking scientists around the world are now sharing their lab notebooks online
It’s never too late to take another look at your data. The incredibly knowledgeable Sally Cowley suggested that I shouldn’t just think about how many cells contain any AlexFluor488 antibody after electroporation, but how bright each cell is (i.e. the amplitude of fluorescence). Looking at this instead will give me an idea of how much Read More …
Background: Scientists centred around University of Oxford came together in the current COVID-19 pandemic to contribute to relevant studies collaboratively. Researchers with no prior experience in virology studies, myself included, had volunteered our time to contribute whichever way we can based on our field of expertise. The Biotech team of SGC Oxford, headed by Nicola Read More …
Background: The binding potency of M4K compounds to ALK2 has been assayed in biochemical assay and cellular assays in HEK293 or C2C12 myoblast cell lines. However, the potency of ALK2 inhibition by M4K compounds has not determined directly in DIPG patient-derived cell lines. While no major deviation from existing assay data is expected, direct experimental Read More …
I decided to test a set of my ALK2 GS loop phosphomutants for activity. As I’ve written about previously, I needed to optimise the concentrations of receptors and decided that 60uM was the right concentration to use as showing activity with the mutants and also being workable in the assay. I set up three different Read More …
So I’m still ‘trying to catch them all’ as the young folks say (!) and building up my collection of purified GS loop mutations so that I can test them all together and start building up a picture of what phosphorylation sites are necessary for SMAD1 activation and which are not. This time around I Read More …
I’ve been working recently on purifying some proteins for use in future experiments and wanted to explain a little bit about what I need each construct for as they’re all for different things. Purification, as always, is a time consuming business but vital for any biophysical experiment that I do given all my work is Read More …
Hello folks, another day another deposition. That makes it sound horribly commonplace but actually Ros put in immense effort into crystallising and solving these proteins and it’s my job just to add the final polish (fix those last stubborn geometry errors), give it a once over to make sure there’s nothing obviously wrong, and whip Read More …
One of the big aims of the M4K project was to look at the structure of potentially interesting molecules bound to the protein ALK2 and use that to try and design better molecules that might be useful in the treatment of the rare childhood brain cancer, DIPG. Ros did a lot of work on this Read More …
I’ve been getting my stocks in order recently and catching up on some other work which has been both busy but not the most thrilling in terms of experimental results. As such I’ve not had much time to write up a blog post so I’ll try and get back on top of that although it’s Read More …
Background: I have been characterising the cellular potency of many M4K compounds against ALK2 and ALK5. Compounds that targets ALK2 and not ALK5 should be further evaluated for their ability to reduce the viability of cells derived from DIPG patients. Although these cultured cells might not reflect what actually happens in a DIPG patient’s brain, Read More …
