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Background: Aside from determining the optimal numbers of DIPG cells to be seeded for viability assay, it is also important to validate that measurements using CellTiter Glo are in agreement with other methods of determining cell viability. Experimental design: I chose to compare the EC50 values of M4K2009 on HSJD-DIPG-007 when measured using different methods. Read More …
As soon as possible I’m hoping to use CRISPR/Cas9 genetic editing to make sets of DIPG patient cells that differ only in one gene (either with/without the whole gene or with/without mutations of interest). This will allow me to make far more accurate comparisons of behaviour between the cells in relation to specific genes or Read More …
Sorry I’ve not written for a while. I had some annual leave to use up and then I’ve been helping out with a residential EMBO course that was being co-organised by one of the group leaders in my department. The course was an introduction to a wide variety of biochemical and biophysical techniques. I was Read More …
Background: Evaluation of the efficacy of M4K compounds in DIPG patient-derived cell lines is essential before any promising compounds can be further tested in mouse xenograft models. This approach can aid in narrowing down clinical compound candidates and reduce the time, resources and animal sacrifice needed downstream. A robust and efficient readout for the changes Read More …
Continuing with our goal to synthesize three compounds as ALK2 inhibitors, let’s refer back to Scheme 2 in the previous post on this topic, compounds 6, 8 and 9 were synthesized in good yields (65-90%), however compound 10 could not be obtained. Different conditions were tested to induce the cyclization of the bromo-pyrazine-2-carboxamide 9 Read More …
Following up on the crystals from my last post I sent them along on the last Diamond trip we had to beamline I04 but regrettably didn’t get any usable datasets. Perhaps fortunately there is a possible explanation for this that isn’t just ‘the crystals were a bit too rubbish’. We send our crystals to the Diamond Read More …
Stem cells are a set of cells that, unlike most cells in the body, can divide as many times as they want, and can turn into any cell in the body. The mutations in cancer often make the cells behave more like stem cells, because in order to grow into a large tumour they may Read More …
I’ve been lucky enough to have a student (Mira) in the lab working with me for the past few weeks and so I’d like to write about the work that we’ve been doing together. We spent some time cleaning up an old sample of ALK2 that Ros had left over in the freezer to set Read More …
I’ve got a few things to catch up with on the blog – one of them is to talk about a conference I attended in Oxford recently. I attended the Biochemical Society BMP Signalling in Cancer II meeting which was held in St. Annes College, Oxford. At this meeting I presented a poster on the Read More …
Special thanks to: David Drewry – Helped with designing the nanoBRET tracers M4K pharma OICR chemist team – Synthesised the M4K1046-linker derivatives Carrow Wells (UNC) – Conjugated the M4K1046 to nanoBRET fluorophore Background: I have always wanted to establish nanoBRET target engagement assay for ALK5. Relative to dual luciferase promoter assay and immunofluorescent staining, nanoBRET Read More …
