Name: | Chicken polyclonal antibody to tyrosine hydroxylase |
Immunogen: | Full length human TH as expressed in and purified from E. coli. |
HGNC Name: | TH |
UniProt: | P07101 |
Molecular Weight: | ~58kDa |
Host: | Chicken |
Isotype: | |
Species Cross-Reactivity: | Human, rat, mouse |
RRID: | AB_2737416 |
Format: | Affinity purified antibody at 1mg/mL in 50% PBS, 50% glycerol plus 5mM NaN3 |
Applications: | WB, IF/ICC, IHC, FACS |
Recommended Dilutions: | WB: 1:50,000. IF/ICC and IHC: 1:10,000 |
Storage: | Stable at 4°C for at least one year. |
Chicken Polyclonal Antibody to Tyrosine Hydroxylase
Cat# CPCA-TH
$150.00 – $1,000.00
Tyrosine hydroxylase (TH) is a vital enzyme responsible for the generation of l-3,4-dihydroxyphenylalanine (L-DOPA) from the amino acid tyrosine. L-DOPA is the direct precursor of the neurotransmitter dopamine, and dopamine can itself be processed to produce the neurotransmitters adrenalin and noradrenalin (a.k.a. epinephrin and norepinephrin respectively). Neurons which use dopamine, adrenalin or noradrenaline, called collectively chatecholamines, must express TH. TH has a very restricted distribution in the brain but is highly expressed in the cells in which it is found. As a result antibodies to TH are useful for the identification of chatecholaminergic neurons. TH positive neurons in the rat are localized into clusters of cells most of which are in the brain stem, including notably the substantia nigra and locus ceruleus (1,2). The clusters of cells are usually referred to by a classification scheme based on that proposed by Dahlstrӧm and Fuxe, which labels cells in groups A1 – A17 and C1 to C3 (2). Subpopulations of neurons are localized in the olfactory bulb, habenula and retina. TH positive cells are also found in a subset of cells in the adrenal medulla, sympathetic ganglia, sensory ganglia and enteric ganglia (2). Numerous TH positive axons can be seen coursing through the striatum and to a much lesser degree the cortex originating from the mid brain A8, A9 and A10 nuclei. TH neurons have a huge impact on brain function and behavior but are relatively infrequent- the rat brain contains about 22,000 TH positive neurons in the A8, A9 and A10 nuclei out of a total of 200 million neurons (3). Parkinson’s disease is caused by the loss of TH positive dopaminergic neurons in the substantia nigra, which are also relatively low in number (4), and perturbation of TH neurons has been implicated in Alzheimer’s disease and schizophrenia (5-7). There is one mammalian gene which produces one mRNA transcript and one protein in rat but four alternate mRNA transcripts produce four slightly different forms of TH proteins in humans (8).
CPCA-TH was made against full length recombinant human TH based on the 524 amino acid sequence in NP_954987.2, expressed in and purified from E. coli. The antibody has an extremely high titre and can be used to study TH positive cells in culture and in sectioned material. The antibody has also been used to efficiently isolate TH positive PBMCs by FACS (9). We also supply a mouse monoclonal and a chicken polyclonal antibodies to this protein, MCA-4H2 and RPCA-TH. Mouse select image at left for larger view.
Chromogenic immunostaining of a formalin fixed paraffin embedded rat brain stem sagittal section with chicken pAb to tyrosine hydroxylase, CPCA-TH, dilution 1:10,000, detected in DAB (brown) following the The sample was processed using our standard IHC protocol outlined here. Hematoxylin (blue) was used as the counterstain. The tyrosine hydroxylase antibody stains perikarya and processes of certain chatecholaminergic neuronal cells. Mouse select image for larger view.
Affinity purified CPCA-TH conjugated with ALEXA647 reliably detects TH expressed in murine peripheral blood mononuclear cells (PBMC). Whole blood acquired from C57B6/J mice via cardiac puncture was diluted 1:1 in sterile PBS and layered on top of Ficoll-Histopaque (GE Healthcare, 17-1440-03) and centrifuged at 400g, room temperature, for 20 minutes with acceleration set to minimum and brakes off. Cells acquired from interface of Ficoll and whole blood were washed twice in sterile PBS, and immediately fixed on ice using the eBioscience Fixation/Permeablization kit (eBioscience, 88-8824-00) for 30 minutes, then washed twice with 1mL permeabilization buffer. Intracellular staining for TH was accomplished by addition of 2μL of affinity purified CPCA-TH directly conjugated to Alexa647 in 100uL staining volume (1:50 dilution from stock antibody conjugate solution at 2.2mg/mL) and incubated at room temperature for 30 minutes. Following two washes in 1mL permeabilization buffer, data were immediately acquired on a Sony SP6800 Spectral Analyzer, and analyzed in a FCS Express 7 software (DeNovo) in the University of Florida ICBR cytometry lab. Data provided by Adithya Gopinath working in the lab of Habibeh Khoshbouei, University of Florida
Clear separation of the positive sample (red) from negative control (black, chicken IgY plus anti-chicken Alexa647 conjugate) indicates clear staining and specificity of chicken anti-TH-647 for TH expressed in murine PBMCs, and shows suitability for flow cytometric analysis of murine PBMCs for TH expression. Further details about this interesting approach are found in Gopinath et al. (2020).
1. Pickel VM, et al. Cellular localization of tyrosine hydroxylase by immunohistochemistry. J. Histochem. Cytochem. 23:1-12 (1975).
2. Bjorklund A, Dunnett SB. Dopamine neuron systems in the brain: an update. Trends Neurosci. 30:194-202 (2007).
3. German DC, Manaye KF. Midbrain dopaminergic neurons (nuclei A8, A9, and A10): three-dimensional reconstruction in the rat. J. Comp. Neurol. 331:297-309 (1993).
4. Daubner SC, Le T, Wang S. Tyrosine hydroxylase and regulation of dopamine synthesis. Arch. Biochem. Biophys. 508:1-12 (2011).
5. Haavik J, Toska K. Tyrosine hydroxylase and Parkinson’s disease. Mol. Neurobiol. 16:285-309 (1988).
6. Torack RM, Morris C. Tyrosine hydroxylase-like (TH) immunoreactivity in Parkinson’s disease and Alzheimer’s disease. . 4:165-71 (1992).
7. Benes FM, Todtenkopf MS, Taylor JB. Differential distribution of tyrosine hydroxylase fibers on small and large neurons in layer II of anterior cingulate cortex of schizophrenic brain. Synapse 25:80-92 (1997).
8. Lewis DA, Melchitzky DS, Haycock JW. Four isoforms of tyrosine hydroxylase are expressed in human brain. Neuroscience 54:477-92 (1993)
9. Gopinath A, et al. J Immunol Methods doi:10.1016/j.jim.2019.112686 476:112686 (2020).
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Contact info
EnCor Biotechnology Inc.
4949 SW 41st Boulevard, Ste 40
Gainesville
Florida 32608 USA
Phone: (352) 372 7022
Fax: (352) 372 7066
E-mail: admin@encorbio.com
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