Characterization of 3067 and 3068 antibodies. A) The top schematic (in blue) shows the domain structure of SNAP25; SN1 and SN2 denote the respective SNARE domains. An alignment of the peptide sequences from rat SNAP25a and SNAP25b that were used for antibody production is also shown. The sequence of rat SNAP23 in the corresponding region is provided for comparison. Numbers indicate the position of the peptides in relation to the full-length proteins. The shading highlights conserved residues in SNAP25a and SNAP25b. The shading for SNAP23 highlights amino acids that are identical to those in the corresponding region of SNAP25b. B) HEK293T cells were transfected with plasmids encoding EGFP-tagged SNAP25a, SNAP25b and SNAP23. Equal volumes of lysates from the transfected cells were probed with antibodies against SNAP25a (3067), SNAP25b (3068), SNAP23 (Synaptic Systems), and GFP (Roche). C) Cerebellum lysates from wild-type or SNAP25a/a mice were resolved by SDS-PAGE and transferred to nitrocellulose for immunoblotting analysis using 3067, 3068, pan-SNAP25 and SNAP23 antibodies. The position of molecular weight markers are shown on the left side of panels B and C.