Mouse Anti-Tyrosine Hydroxylase (TH, Tyrosine 3-Hydroxylase, Tyrosine 3-Monooxygenase)
Tyrosine hydroxylase is an excellent marker for dopaminergic and noradrenergic neurons. Tyrosine hydroxylase (a.k.a. tyrosine 3-monooxygenase) is the enzyme responsible for catalyzing the conversion of the amino acid L-tyrosine to L-3,4-dihydroxyphenylalanine (L-DOPA). L-DOPA is a precursor for dopamine, which, in turn, is a precursor for the important neurotransmitters norepinephrine (noradrenaline) and epinephrine (adrenaline). Tyrosine hydroxylase catalyzes the rate limiting step in this synthesis of catecholamines. In humans, tyrosine hydroxylase is encoded by the TH gene, and the enzyme is present in the central nervous system (CNS), peripheral symphatic neurons and the adrenal medulla. The enzymatic activity of TH requires ferrous ions as cofactors and is believed to be regulated by phosphorylation. At least four isoforms of human TH have been identified which result from alternative splicing. Tyrosine hydroxylase, phenylalanine hydroxylase and tryptophan hydroxylase together make up the family of aromatic amino acid hydroxylases (AAAHs).
Applications
Suitable for use in Immunofluorescence, Flow Cytometry, Western Blot, Immunoprecipitation, Immunohistochemistry. Other applications not tested.
Recommended Dilution
Flow Cytometry: Fixed, permeabilized dopaminergic nerve terminals from rat striatum. (See: Wolf, ME, Kapatos, G. 1989. The Journal of Neuroscience, January 1989, 9(l): 108-114; Wolf ME, Zigmond, MJ, Kapatos, G .1989. J. Neurochemistry 53(3):879-885). Immunofluorescence: 1:200-1:1000. 4% PFA fixed tissues/cells permeabilized with 0.1-0.4% triton X-100. Also works in fresh frozen and acetone-fixed tissues/cells. Western Blot: 1:1000-1:2000 using SDS-reduced samples. Detects a 59-61kD protein. Rat brain lysate is a suitable control. Immunohistochemistry(FFPE): 1:100-1:200, depending on incubation time and detection method used. Requires heat-induced epitope retrieval. Immunoprecipitation: 1:100, immobilized on protein A beads (see Fleming-Jones, et al. 1995. J. Protein Chemistry. 14(5):275-282) Optimal dilutions to be determined by the researcher
Storage and Stability
Lyophilized and reconstituted products are stable for 12 months after receipt at -20°C. Reconstitute with sterile ddH2O. Aliquot to avoid repeated freezing and thawing. Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Immunogen
PC12 cells. Epitope: located on the outside of the regulatory N-terminus
Form
Supplied as a lyophilized powder. Reconstitute in 100ul sterile ddH2O.
Specificity
Recognizes rat Tyrosine Hydroxylase. Does not react with dopamine-beta-hydroxylase, phenylalanine hydroxylase, trytophan hydroxylase, dehydropteridine reductase, sepiapterin reductase or phenethanolamine-N-methyl transferase (PNMT). Species Crossreactivity: chicken, frog, equine, human, monkey, mouse, vole, sheep, zebrafish
