Anti-Tyrosine Hydroxylase antibody (ab6211)

Tyrosine hydroxylase antibody (ab6211) staining rat mesencephalic serotoninergic neurons. Pictures taken using objectives X20 (A) or X20 (B). Note that the staining is observed in the cell body and the processes of these neurons. ab6211 was used at 1/1000 incubated for 2 days at room temperature. Secondary antibody was anti-rabbit Alexa Fluor 488 used at 1/1000 for 2 hours at room temperature. Rat brain tissue (ab29475) was perfusion fixed (paraformaldehyde 4% ) followed by post-fixation overnight in the same fixative, cryoprotected in 20% sucrose and then frozen in OCT. 30µm coronal sections of brain were cutting by cyrostat and immunohistochemistry was performed as the 'free floating' technique.

ab6211 staining tyrosine hydroxylase in PC-12 cells treated with orexin A (ab120212), by ICC/IF. Decrease in expression of tyrosine hydroxylase correlates with increased concentration of orexin A, as described in literature.
The cells were incubated at 37°C for 1h in media containing different concentrations of ab120212 (orexin A) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum (ab7481), 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab6211 (1/1000 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

Three bands are detected at ≈ 42 kDa, 53 kDa and 84 kDa.

Immunohistochemical detection of dopaminergic neurons in the rat zona incerta in a formalin-fixed floated cryostat section. Tyrosine hydroxylase immunoreactivity was visualized with ab6211 (1:100,000), using the biotinylated secondary antibody-ABC method and nickel-diaminobenzidine chromogen. Photo courtesy of Dr. Erik Hrabovszky, Hungarian Academy of Sciences, Budapest, Hungary.