Skip to main content
[Preprint]. 2024 Sep 27:2024.09.25.614766. [Version 1] doi: 10.1101/2024.09.25.614766

Figure 3. IP-FCM HTT detection assay signal is influenced by protein fragmentation, the position of the affinity tag and the oligomerization state of the protein.

Figure 3.

A. MW1-HDB4 IP-FCM analysis of full-length (FL) HTT, fusion HTT Q68 and N586 HTT Q68 protein with approximately the same Q-length. B. HDB4/MW1 IP-FCM analysis of full-length HTT with polyQ tracts spanning either 23 or 66 glutamines, with N or C-terminal FLAG-tag. C. Left – Gel filtration (GF) trace of FLAG-affinity chromatography purified full-length HTT Q54 applied to Superose6 10/300 GL column which elutes across fractions 1-8 (F1-F8). Middle – SDS-PAGE analysis of FLAG-affinity chromatography flow through, wash, elution and GF input fractions. Right - HDB4/MW1 IP-FCM analysis of concentration normalized GF fractions F1-F8 at different dilutions. IP-FCM graphs shown are generated from a representative replicate dataset, N=3.