Mouse Anti-PolyQ Tract Recombinant Antibody (clone 1C2) (CAT#: PABL-307)

Figure 1 GST pull-down assays of 3B5H10 and 1C2 on GST-polyQ bearing different polyQ lengths: 3B5H10 and 1C2 come down in the presence of GST-Q11, GST-Q22 or GST-Q41 but not GST.

Klein, F. A., Zeder-Lutz, G., Cousido-Siah, A., Mitschler, A., Katz, A., Eberling, P.,... & Trottier, Y. (2013). Linear and extended: a common polyglutamine conformation recognized by the three antibodies MW1, 1C2 and 3B5H10. Human molecular genetics, 22(20), 4215-4223.

Figure 2 Interaction of 3B5H10-Fab and 1C2-Fab with polyQ of various lengths as analyzed by WB.

The efficiency of detection of polyQ stretches by WB, employing 1C2-Fab or 3B5H10-Fab as primary antibodies, increases in a polyQ-length-dependent manner. Equal amounts of GST-Q11, GST-Q22 and GST-Q41 were mixed and loaded in a unique well. After SDS–PAGE and transfer onto a nitrocellulose membrane, the membrane was cut into 15 equivalent lanes. Each lane was individually incubated with the following primary antibodies, at the indicated concentrations: 1C2 IgG (lane 1); 1C2-Fab (lanes 2 to 7); anti-GST (lane 8); 3B5H10-Fab (lanes 9–14); 3B5H10 IgG (lane 15). The 15 lanes were then reassembled, incubated with GAM-peroxidase and revealed simultaneously for 30 s (top panel) or 1110 s (lower panel). Note the equal amounts of the three GST-polyQ proteins as revealed with anti-GST after a 30 s exposure.

Klein, F. A., Zeder-Lutz, G., Cousido-Siah, A., Mitschler, A., Katz, A., Eberling, P.,... & Trottier, Y. (2013). Linear and extended: a common polyglutamine conformation recognized by the three antibodies MW1, 1C2 and 3B5H10. Human molecular genetics, 22(20), 4215-4223.

Figure 3 Inhibition of HD exon 1 fibrillogenesis by antibodies.

(A) Effect of increasing concentrations of 1C2 antibody or mouse IgG 2a on mycHD51 aggregation. Aggregate formation was detected by the filter retardation assay. (B) Quantitative analysis of dot-blot results shown in A. The relative amount of aggregate for each sample was quantified on a Fuji-Imager (LAS 2000). For each experiment, the signal intensity obtained from the samples without added antibodies was arbitrarily set as 100.

Heiser, V., Scherzinger, E., Boeddrich, A., Nordhoff, E., Lurz, R., Schugardt, N.,... & Wanker, E. E. (2000). Inhibition of huntingtin fibrillogenesis by specific antibodies and small molecules: implications for Huntington's disease therapy.Proceedings of the National Academy of Sciences, 97(12), 6739-6744.

Figure 4 Soluble Htt and formic acid-dissolved and non-dissolved (arrow) Htt aggregates were detected on Western blot by the 1C2 antibody.

A, soluble and insoluble fractionation of Neuro-2a cell lysate after transient transfection of Htt-exon1-97Q or an empty vector as control. Six hours after transfection, cells were treated for 16 h with DMSO, epoxomicin, or 3-MA and harvested. Soluble Htt and formic acid-dissolved and non-dissolved (arrow) Htt aggregates were detected on Western blot by the 1C2 antibody. SDS-insoluble Htt aggregates were analyzed by a filter trap assay in doublets. β-Actin was used as a loading control.

Juenemann, K., Schipper-Krom, S., Wiemhoefer, A., Kloss, A., Sanz, A. S., & Reits, E. A. (2013). Expanded polyglutamine-containing N-terminal huntingtin fragments are entirely degraded by mammalian proteasomes. Journal of Biological Chemistry, 288(38), 27068-27084.