Anti-EB3 antibody [KT36] (ab53360) at 1 µg/ml + Human brain tissue lysate - total protein (ab29466) at 10 µg

Secondary
Peroxidase Conjugated AffiniPure Rabbit Anti-Rat IgG (H+L) at 1/10000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 32 kDa
Observed band size: 32 kDa


Exposure time: 8 minutes

Immunofluorescent staining for COS7 cells overexpressing EB3, using ab53360 at 1/100.

Immunofluorescent staining of cell mitosis using ab53360 crude culture supernatant diluted 1:10 (Please note that there was about 50-100 ug/ml antibody in the supernatants) COS7 Transformed African Green Monkey Kidney Fibroblast Cells.

Overlay histogram showing SH-SY5Y cells stained with ab53360 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab53360, 2µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgG2a [aRTK2758] (2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.