The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. Positive result against immunising peptide.
1/1000. Detects a band of approximately 52 kDa (predicted molecular weight: 55.4 kDa).Can be blocked with GABA A Receptor alpha 5 peptide (ab137922). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
Use at an assay dependent concentration.
Use a concentration of 5 µg/ml.
GABA, the major inhibitory neurotransmitter in the vertebrate brain, mediates neuronal inhibition by binding to the GABA/benzodiazepine receptor and opening an integral chloride channel.
Belongs to the ligand-gated ion channel (TC 1.A.9) family. Gamma-aminobutyric acid receptor (TC 1.A.9.5) subfamily. GABRA5 sub-subfamily.
Immunocytochemistry/ Immunofluorescence - Anti-GABA A Receptor alpha 5 antibody (ab10098)
ICC/IF image of ab10098 stained SHSY5Y cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab10098, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Immunohistochemistry (Frozen sections) - Anti-GABA A Receptor alpha 5 antibody (ab10098)Image courtesy of Tara Stewart by Abreview.
ab10098 staining GABA A Receptor alpha 5 in rat hippocampus sections by Immunohistochemistry (Frozen sections). Rats were transcardially perfused with 4% PFA, the brain was cryoprotected in 30% sucrose before flash freezing. Coronal sections were taken using cryostat (20 micron sections) and mounted onto slides. Tissue was fixed in paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were then blocked using 10% serum for 1 hour at 20°C and then incubated with ab10098 at a 1/200 dilution for 16 hours at 4°C. The secondary used was an Alexa Fluor® 594 goat anti-rabbit IgG (H+L) used at a 1/600 dilution. An antigen retrieval step improved results (5% SDS for 5 minutes before standard IHC protocol).