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Rabbit anti-Tyrosine Hydroxylase (Catalog Number AB152) localization of tyrosine hydroxylase in culture of primary neurons. Photo courtesy of Dr. Mehd ...
anti-Tyrosine Hydroxylase (TH) staining in mouse primary neural cultures using AB152 shown with an FITC fluorescent secondary (green). Fixation was 4 ...
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Anti-Tyrosine Hydroxylase
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| Dr, Ft, H, M, R, Fe, Ml, Sqd | ELISA, IP, WB, IH, IH(P) | Rabbit | Affinity Purified | Polyclonal Antibody |
UniProt Number:
UniProt Summary
FUNCTION: SwissProt: P07101 # Plays an important role in the physiology of adrenergic neurons.
COFACTOR: Fe(2+) ion.
SIZE: 528 amino acids; 58524 Da
TISSUE SPECIFICITY: Mainly expressed in the brain and adrenal glands.
DISEASE: SwissProt: P07101 # Defects in TH are the cause of autosomal recessive Segawa syndrome [MIM:605407]; also known as DOPA-responsive dystonia. Typically, it begins in childhood or adolescence with progressive difficult ... see more »
COFACTOR: Fe(2+) ion.
SIZE: 528 amino acids; 58524 Da
TISSUE SPECIFICITY: Mainly expressed in the brain and adrenal glands.
DISEASE: SwissProt: P07101 # Defects in TH are the cause of autosomal recessive Segawa syndrome [MIM:605407]; also known as DOPA-responsive dystonia. Typically, it begins in childhood or adolescence with progressive difficult ... see more »
Entrez Gene Number:
Entrez Gene Summary
Tyrosine hydroxylase is involved in the conversion of tyrosine to dopamine. As the rate-limiting enzyme in the synthesis of catecholamines, tyrosine hydroxylase has a key role in the physiology of adrenergic neurons.
Description:
Anti-Tyrosine Hydroxylase
Background Information:
Tyrosine hydroxylase plays an important role in the physiology of adrenergic neurons. It is the first and rate-limiting enzyme involved in the biosynthesis of the catecholamines Dopamine and Norepinephrine from tyrosine. TH is, therefore, a useful marker for dopaminergic and noradrenergic neurons. 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.
Alternate Names:
TH; Tyrosine Monooxygenase
Specificity:
Recognizes Tyrosine hydroxylase. It is expected that the antibody will react with most mammalian and many non-mammalian species. It has been reported that this antibody does not work on paraffin embedded human tissue.
Key Applications:
- ELISA
- Immunoprecipitation
- Western Blotting
- Immunohistochemistry
- Immunohistochemistry (Paraffin)
Application Notes:
Immunohistochemistry: 1:100-1:500, 4°C 1-3 days. Stains both fresh frozen and paraffin embedded tissue samples by indirect immunofluorescence and immunoperoxidase.
Western blot. 0.1-1µg/mL. Detects a band at approximately 60 kDa corresponding to tyrosine hydroxylase.
Immunoprecipitation
Optimal dilutions must be determined by the end user.
GENERAL METHODOLOGY FOR RAT BRAIN PERFUSION AND STAINING USING AB152
1. Perfuse the animal with PBS with 4% paraformaldehyde fixative. (Glutaraldehyde [0.1-0.5%] may be used.)
2. Remove brain and place it in cold fixative without glutaraldehyde (2-4 hours postfixation at 0-4°C).
3. Transfer the brain (or blocks therefrom) to PBS containing at least 5% sucrose. When ready to section, place the tissue on cryostat chucks and freeze them on crushed dry ice. Alternatively, freeze the sample on the stage of a sliding microtome or glue the sample onto a Vibratome chuck.
4. Make 15-30 micron sections and place them in PBS. Rinse the sections in PBS several times.
5. Place the sections in blocking solution (PBS containing 10% goat serum, 1% RIA-BSA, 0.3% Triton X-100 [high purity]) for 30-60 minutes at room temperature.
6. After a brief rinse in carrier solution (optional), place the sections in primary antibody that has been diluted in carrier solutions (PBS containing 1% goat serum, 1% RIA-BSA, 0.3% Triton X-100 [high purity]). Final working dilutions to be determined by end user. Suggested starting dilution is 1:100-1:500 of the antibody. Successful incubation conditions include 1-3 days at 0-4°C, 2-18 hours at room temperature, and 1-4 hours at 37°C.
7. Rinse the sections 3x with the carrier solution and place the sections in secondary antibody reagent. This is usually labeled or unlabeled goat anti-rabbit antibody, depending upon the visualization method to be used. For peroxidase reactions, the typical incubation is 1 hour at 37°C or 2 hours at room temperature.
8. Rinse the sections 3x with the carrier solution. For fluorescence labeling, rinse an additional time with PBS alone and mount from distilled water.
9. For peroxidase-anti-peroxidase reactions, place the sections in carrier solution containing the tertiary antibody (substitute Tween 20 or Nonidet P40 for Triton X-100). For avidin-biotin-complex reactions, place the sections in PBS containing either Tween 20 or Nonidet P40. Incubate the sections for 1-2 hours at room temperature and rinse them 3x in PBS alone.
10. For peroxidase development, place the sections in PBS containing diaminobenzide (50 mg/100 mL) and add 33 µL of 30% H2O2. After 2-30 minutes at room temperature, rinse the sections well with PBS, mount the sections from dilute PBS (e.g. 1:5, PBS:H2O), and air dry. Rinse the slides in several changes of distilled water, dehydrate and clear.
Western blot. 0.1-1µg/mL. Detects a band at approximately 60 kDa corresponding to tyrosine hydroxylase.
Immunoprecipitation
Optimal dilutions must be determined by the end user.
GENERAL METHODOLOGY FOR RAT BRAIN PERFUSION AND STAINING USING AB152
1. Perfuse the animal with PBS with 4% paraformaldehyde fixative. (Glutaraldehyde [0.1-0.5%] may be used.)
2. Remove brain and place it in cold fixative without glutaraldehyde (2-4 hours postfixation at 0-4°C).
3. Transfer the brain (or blocks therefrom) to PBS containing at least 5% sucrose. When ready to section, place the tissue on cryostat chucks and freeze them on crushed dry ice. Alternatively, freeze the sample on the stage of a sliding microtome or glue the sample onto a Vibratome chuck.
4. Make 15-30 micron sections and place them in PBS. Rinse the sections in PBS several times.
5. Place the sections in blocking solution (PBS containing 10% goat serum, 1% RIA-BSA, 0.3% Triton X-100 [high purity]) for 30-60 minutes at room temperature.
6. After a brief rinse in carrier solution (optional), place the sections in primary antibody that has been diluted in carrier solutions (PBS containing 1% goat serum, 1% RIA-BSA, 0.3% Triton X-100 [high purity]). Final working dilutions to be determined by end user. Suggested starting dilution is 1:100-1:500 of the antibody. Successful incubation conditions include 1-3 days at 0-4°C, 2-18 hours at room temperature, and 1-4 hours at 37°C.
7. Rinse the sections 3x with the carrier solution and place the sections in secondary antibody reagent. This is usually labeled or unlabeled goat anti-rabbit antibody, depending upon the visualization method to be used. For peroxidase reactions, the typical incubation is 1 hour at 37°C or 2 hours at room temperature.
8. Rinse the sections 3x with the carrier solution. For fluorescence labeling, rinse an additional time with PBS alone and mount from distilled water.
9. For peroxidase-anti-peroxidase reactions, place the sections in carrier solution containing the tertiary antibody (substitute Tween 20 or Nonidet P40 for Triton X-100). For avidin-biotin-complex reactions, place the sections in PBS containing either Tween 20 or Nonidet P40. Incubate the sections for 1-2 hours at room temperature and rinse them 3x in PBS alone.
10. For peroxidase development, place the sections in PBS containing diaminobenzide (50 mg/100 mL) and add 33 µL of 30% H2O2. After 2-30 minutes at room temperature, rinse the sections well with PBS, mount the sections from dilute PBS (e.g. 1:5, PBS:H2O), and air dry. Rinse the slides in several changes of distilled water, dehydrate and clear.
Species Reactivity:
- Drosophila
- Ferret
- Human
- Mouse
- Rat
- Feline
- Mollusk
- Squid
Immunogen:
Denatured tyrosine hydroxylase from rat pheochromocytoma (denatured by sodium dodecyl sulfate).
Molecular Weight:
62 kDa
Control:
Positive control: Brain (corpus striatum, sympathetic nerve terminals) and adrenal glands
Negative Control: Liver
Negative Control: Liver
Presentation:
Affinity Purified immunoglobulin. Liquid in 10 mM HEPES (pH 7.5), 150 mM NaCl (pH 7.5), 100 mg/mL BSA and 50% glycerol.
Storage Conditions:
Maintain frozen at -20 to -70°C in undiluted aliquots for up to 6 months after date of receipt. Avoid repeated freeze/thaw cycles. Do not store in a self defrosting freezer.
Gene Symbol:
- TH
- TYH
- EC 1.14.16.2
Antibody Category:
Neuroscience
Antibody Sub-Category:
- Neurotransmitters & Receptors
- Neuronal & Glial Markers
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Descriptive Text:
Tyrosine Hydroxylase catalyzes the rate-limiting step in the production of Dopamine.
Concentration:
0.08 mg/mL
Format Code:
APur
Trade Name:
Chemicon (Millipore)
Purification Method:
Immunoaffinity Purified
Format:
Affinity Purified
Antibody Type:
Polyclonal Antibody
Host:
Rabbit