Recombinant Anti-Tyrosine Hydroxylase antibody [EP1533Y] (ab75875)

Immunohistochemical analysis of rat brain tissue section labeling Tyrosine Hydroxylase with ab75875 at 1/1000 dilution. The tissue was fixed with Formaldehyde. Antigen retrieval was heat mediated using citric acid. A biotin conjugated goat anti-rabbit polyclonal IgG at 1/250 was used as the secondary antibody.

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ab75875 staining Tyrosine Hydroxylase in Pig Myocardiun tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/1000 in blocking buffer) for 2 hours at 21°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.

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IHC image of Tyrosine Hydroxylase staining in human cerebral cortex formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab75875, 1/200 dilution, 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.

ICC/IF image of ab75875 stained SKNSH 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 ab75875 at 1/100 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.

Immunohistochemical analysis of mouse brain tissue section labeling Tyrosine Hydroxylase with ab75875 at 1/800 dilution. The tissue was fixed with Formaldehyde. Antigen retrieval was heat mediated using citric acid. A biotin conjugated goat anti-rabbit polyclonal IgG at 1/250 was used as the secondary antibody.

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