
Month: July 2017
Huntingtin and huntingtin-interacting protein purification 2017/07/21
This past week I have been working hard to make lots of protein for future experiments. I have successfully purified full-length C-terminally tagged huntingtin. Having the FLAG tag on the C-terminal end of the protein seems to reduce the aggregation of the sample – I have now seen this consistently in 2 different purifications. I Read More …
Purification of high mobility group box 1 protein (HMGB1) – a huntingtin interacting protein
To generate better data for cryo-EM analysis of the huntingtin protein structure, more stable and conformationally homogeneous samples may help improve the particle averages. Purifying complexes of huntingtin with proteins which specifically bind huntingtin may promote such sample stability. High mobility group box 1 (HMGB1) is known to bind huntingtin but, to my knowledge, this interaction Read More …
Purifying huntingtin with a C-terminal FLAG tag from insect cells
Nearly all of my experiments, whether interaction or structural studies, require large (ish) amounts of purified huntingtin protein. This is why a lot of my posts and write ups describe making the huntingtin protein. Over a year into this project, I have now established good protocols for expressing and purifying the protein in large quantities Read More …