Immunohistochemical staining of aquaporin 1 in rat brain.

Overlay histogram showing HEK293 cells stained with ab9566 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Triton 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 (ab9566, 2µg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Triton used under the same conditions.

Immunohistochemical analysis of exocrine pancreas cells labeling Aquaporin 1 with ab9566. 

Anti-Aquaporin 1 antibody [1/22] (ab9566) at 1 µg/ml + Human skeletal muscle tissue lysate - total protein (ab29330) at 10 µg

Secondary
Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 29 kDa
Observed band size: 29 kDa
Additional bands at: 50 kDa, 75 kDa. We are unsure as to the identity of these extra bands.


Exposure time: 30 seconds