This past week I have been replenishing my stocks of huntingtin samples making both the full-length Q42 protein as well as the full-length Q23 protein in complex with HAP40. The protocol for HTT-HAP40 complex generation is now very robust and the sample looks top notch.
I have also tried to make HTT complexes with other putative interactors, it has been challenging! Using the same protocol I use to make HTT-HAP40 I looked at PRC2, HDAC6, USP9X amongst other potential huntingtin binding proteins. Unfortunately these experiments did not pan out too well but I now have a clearer idea of the potential technical obstacles as well as a plan for how to proceed forward with the interaction studies.
As always, any ideas, feedback, comments or suggestions are more than welcome!