Recombinant Anti-SNAP25 antibody [EPR3275] (ab109105)

Flow Cytometry analysis of Neuro-2a(Mouse neuroblastoma neuroblast) cells labelling SNAP25 with Purified ab109105 at 1:200 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor™ 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

False colour image of Western blot: Anti-SNAP25 antibody [EPR3275] staining at 1/1000 dilution, shown in green; loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) staining at 1/20000 dilution, shown in red. In Western blot, ab109105 was shown to bind specifically to SNAP25. A band was observed at 27 kDa in wild-type SH-SY5Y cell lysates with no signal observed at this size in SNAP25 knockout cell line ab280041 (knockout cell lysate ab280100). To generate this image, wild-type and SNAP25 knockout SH-SY5Y cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.