GABA, Sigma-Aldrich, A2052

The antibody was used for Immunofluorescence staining of neurons derived from murine ES cells differentiated in culture.
The cells were grown on coated cover slips, fixed for 30min in 4% PFA and treated for 5 min with 0,005% Triton-X-100. To reduce unspecific binding cells are blocked/ quenched with a 1% BSA in PBS solution.
The Antibody was diluted 1:250. The secondary antibody was a goat anti rabbit 586 diluted 1:800.
A Strong staining of the cytoplasm into the neurites was observed. The antibody worked well in double staining experiments with Sigma # T5076 mouse-anti-β-III-tub (1:8000) or bdbiosciences # 612300 mouse-anti-TH (1:500).

We tried this antibody on mouse cortex and thalamus by western blot. There was no clear specific band detected at 1:300.

I have used this antibody for IHC of mouse spinal cord cryo-sections using fluorescent antibodies in order to detect GABAerge neurons. Staining of GABA requires fixation with 4% PFA and 0.2% glutaraldehyde, and subsequent treatment of the tissue with NaBH4 to reduce autofluorescence of glutaraldehyde. The antibody penetrates the tissue very slowly. GABA immunoreactivity is observed in somas as well as in processes.
Dilution: 1:250.

This antibody was tested on tissue and cultured primary hippocampal neurons using fluorescent antibodies. In both cases, the antibody worked very well.
Dilution 1:500