ICC/IF image of ab35971 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 (ab35971, 1µg/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.
IHC image of ab35971 staining in human normal placenta formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH9, epitope retrieval solution 2) for 20 mins. The section was then incubated with ab35971, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-GABA Receptor Epsilon antibody (ab35971)Image from Hengen KB et al., PLoS One. 2012;7(1):e30608. Epub 2012 Jan 31.Fig 6.; doi:10.1371/journal.pone.0030608; January 31, 2012, PLoS ONE 7(1): e30608.
Immunohistochemical analysis of male, virgin female, G17 pregnant, and 30 d post-partum rat medullary neurons, staining GABA Receptor Epsilon with ab35971 at 1/1000 dilution.
Western blot - Anti-GABA Receptor Epsilon antibody (ab35971)
Anti-GABA Receptor Epsilon antibody (ab35971) (0.2-1 ug/ml) + Human Placenta
Fatemi SH et al. Expression of GABAA a2-, ß1- and ?-receptors are altered significantly in the lateral cerebellum of subjects with schizophrenia, major depression and bipolar disorder. Transl Psychiatry3:e303 (2013).
Read more (PubMed: 24022508) »
Hengen KB et al. Increased GABA(A) Receptor e-Subunit Expression on Ventral Respiratory Column Neurons Protects Breathing during Pregnancy. PLoS One7:e30608 (2012).
Read more (PubMed: 22303446) »