Purification and preparation of huntingtin samples for cryo-EM grid preparation in Oxford

This past 3 weeks I have been in the UK, working furiously to try to get a better handle on the EM structure solution pipeline, both in the lab and at the computer.

I was very fortunate to have the opportunity to prepare my huntingtin protein sample in Susan Lea’s lab, with lots of help from Justin Deme. This meant I could apply incredibly freshly prepared protein to the electron microscopy grids for structural analysis, instead of the freeze-thawed sample used previously, as I usually had to ship the sample on dry ice from Toronto to Oxford. It will be interesting to see if this makes a difference to the sample as it has been treated more gently, I certainly wouldn’t liked to be pulled in and out of the -80C freezer!

Huntington purification

Getting to grips with particle picking! The white boxes select single molecules of huntingtin from the image for subsequent analysis.

Here is the write up from the protein purification and gradient fixation steps of this process on Zenodo. More on this soon as I find time in between all of the cracking talks at the CHDI conference to get this completely written up.

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