Figure 1: Characterisation of a novel monoclonal antibody, DTE41, targeting glutamylated Δ2 form of α-tubulin.

(a) Antigen used to generate mAb DTE41. The C-terminal part of Δ2 form with di-glutamylation was used. Colour codes of the rod structure of antigen peptide are green, carbohydrate chain in Δ2 form; yellow, carbohydrate chain of branched di-glutamates; blue, nitrogen; red, oxide. The peptide has in total 8 carboxyl groups. (b) Schematic view of recombinant α-tubulin tail fused to maltose binding protein (MBP) with different C-terminal modifications, tyrosination (Tyr), detyrosination (deTyr), and Δ2 form. Epitope of DM1A is indicated. (c) Preference of mAb DTE41 to Δ2-tubulin. Recombinant α-tubulin C-termini with different modifications were subjected to immunoblotting. Original full-length blot images were provided in Supplementary Figure 6a. (d) Schematic view of peptides used for ELISA. (e) ELISA of mAb DTE41 against peptides shown in panel d.