Anti-GABA A Receptor beta 3/GABRB3 antibody [N87/25] (ab98968) at 1/1000 dilution + Rat brain membrane lysate at 15 µg

Secondary
Sheep Anti-Mouse IgG: HRP

Predicted band size: 55 kDa

Block: 1.5% BSA for 30 minutes at RT.

Incubation with primary antibody for 2 hours at RT.

Incubation with secondary antibody for 1 hour at RT.

ab98968 staining GABA A Receptor beta 3/GABRB3 in mouse brain tissue sections by Immunohistochemistry (IHC-Fr - frozen sections).

Tissue was fixed in 10% formalin for 12-24 hours at RT. Primary Antibody: ab98968 at 1:1000 for 1 hour at RT. Secondary Antibody: HRP/DAB Detection System: Biotinylated Goat Anti-Mouse, Streptavidin Peroxidase, DAB Chromogen (brown) for 30 minutes at RT. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 250-500 µl for 5 minutes at RT.

Overlay histogram showing SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells stained with ab98968 (red line).

The cells were fixed with 4% paraformaldehyde (10 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. The cells were then incubated with the antibody (ab98968, 1µg/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, 2µg/1x10⁶ cells) used under the same conditions.

Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 minutes)/permeabilized with 0.1% PBS-Tween for 20 minutes used under the same conditions.