All lanes : Anti-SYT2 antibody [ZNP-1] (ab154035) at 1 µg/ml

Lane 1 : Human brain tissue lysate - total protein (ab29466)
Lane 2 : Rat Brain Homogenate Lysate
Lane 3 : Mouse Brain Homogenate Lysate
Lane 4 : Zebrafish Brain Homogenate Lysate

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat polyclonal to Mouse IgG – HRP at 1/1000 dilution

Predicted band size: 48 kDa
Observed band size: 65 kDa why is the actual band size different from the predicted?

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebellum tissue sections labeling SYT2 with purified ab154035 at 1/4000 dilution (0.25 µg/mL). Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 minutes. Leica mouse (DS9800) was used as the secondary antibody. Negative control: secondary antibody only control. Hematoxylin was used as a counterstain.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebellum tissue sections labeling SYT2 with purified ab154035 at 1/4000 dilution (0.25 µg/mL). Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 minutes. Leica mouse (DS9800) was used as the secondary antibody. Negative control: secondary antibody only control. Hematoxylin was used as a counterstain.

Immunocytochemistry with SH-SY5Y cells stained with Anti-SYT2 antibody (ab154035) in red and DAPI in blue, as a counter stain. The target protein locates to the plasma membrane (red).

Fixation: 4% Paraformaldehyde, 20 min.
Permeabilization: 0.1% Triton X-100, 10min.
Blocking: 10% Goat Serum/PBS, 1 hour.
Primary: Anti-SYT2 antibody (ab154035) 5 µg/ml in 10% Goat Serum/PBS, 2hr at room temperature or over night at 4°C.
Secondary: Alexa Fluor® 594 Goat Anti-Mouse (H&L) used at a 1/1000 dilution in 10% Goat Serum/PBS, 1hr at RT.
Washing: 3x 1% Goat Serum/PBS, 10 mins/wash
DAPI: 2 µM in 1% Goat Serum/PBS, 10 mins