Anti-Kir2.1/KCNJ2 antibody [N112B/14] (ab85492) at 1/1000 dilution + Monkey COS transient cell lysate at 15 µg with 1.5% BSA for 30 minutes at RT

Secondary
Sheep Anti-Mouse IgG: HRP for 1 hour at RT.

Predicted band size: 48 kDa

ab85492 staining Kir2.1/KCNJ2 in mouse backskin tissue by IHC-P (Bouin's fixed paraffin embedded).

Incubated with primary antibody at 1:1000 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT.

ICC/IF image of ab85492 stained SK-N-SH (Human neuroblastoma cell line) cells.

The cells were 4% formaldehyde fixed (10 minutes) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab85492, 10 µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight^® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1 hour. Alexa Fluor^® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.

ab85492 staining Kir2.1/KCNJ2 in human hippocampal tissue by IHC-P (Bouin's fixed paraffin embedded).

Incubated with primary antibody at 1:1000 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT.

Overlay histogram showing SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells stained with ab85492 (red line).

The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. 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 (ab85492, 1 µg/1x10⁶ cells) for 30 minutes at 22ºC. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2 µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive result in SH-SY5Y cells fixed with 4% paraformaldehyde (10 minutes)/permeabilized in 0.1% PBS-Tween for 20 minutes used under the same conditions.