Human/Mouse Tyrosine Hydroxylase Antibody

(2 Reviews)
  
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human Tyrosine Hydroxylase in direct ELISAs and human, mouse and rat Tyrosine Hydroxylase in Western blots. In direct ELISAs, no cross-reactivity with recombinant human Tryptophan Hydroxylase-1 is observed.
  • Source
    Monoclonal Mouse IgG2B Clone # 779427
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human Tyrosine Hydroxylase
    Ala278-Tyr401
    Accession # P07101
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.25 µg/mL
    See below
  • Simple Western
    5 µg/mL
    See below
  • Immunohistochemistry
    8-25 µg/mL
    See below
  • Immunocytochemistry
    8-25 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples

Detection of Human, Mouse, and Rat Tyrosine Hydroxylase by Western Blot. Western blot shows lysates of human adrenal gland tissue, mouse adrenal gland tissue, and rat adrenal gland tissue. PVDF membrane was probed with 0.25 µg/mL of Mouse Anti-Human/Mouse Tyrosine Hydroxylase Monoclonal Antibody (Catalog # MAB7566) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). Specific bands were detected for Tyrosine Hydroxylase at approximately 50-60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry

Tyrosine Hydroxylase in Mouse Dopaminergic Neurons. Tyrosine Hydroxylase was detected in immersion fixed mouse embryonic stem cells differentiated into dopaminergic neurons using Mouse Anti-Human/Mouse Tyrosine Hydroxylase Monoclonal Antibody (Catalog # MAB7566) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the Northern­Lights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007). Cells were double stained using the Northern­Lights™ 637-conjugated Mouse Anti-Neuron-specific beta-III Tubulin Monoclonal Antibody (white; Catalog # NL1195V). Cells were counterstained with DAPI (blue). Specific staining of Tyrosine Hydroxylase was localized to cytoplasm of dopaminergic neurons. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry

Tyrosine Hydroxylase in Human Brain. Tyrosine Hydroxylase was detected in immersion fixed paraffin-embedded sections of human brain (medulla) using Mouse Anti-Human/Mouse Tyrosine Hydroxylase Monoclonal Antibody (Catalog # MAB7566) at 25 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to neurons. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human and Mouse Tyrosine Hydroxylase by Simple WesternTM. Simple Western lane view shows lysates of human adrenal gland tissue and mouse adrenal gland tissue, loaded at 0.2 mg/mL. Specific bands were detected for Tyrosine Hydroxylase at approximately 52-61 kDa (as indicated) using 5 µg/mL of Mouse Anti-Human/Mouse Tyrosine Hydroxylase Monoclonal Antibody (Catalog # MAB7566). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.5 mg/mL.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Tyrosine Hydroxylase
TH (Tyrosine 3-hydroxylase), also known as Tyrosine 3-monooxygenase, is a 60-62 kDa member of the biopterin-dependent aromatic amino acid hydroxylase family of molecules. It is expressed by neurons of the dopamine and autonomic nervous system, plus the neuroendocrine cells of the adrenal medulla. TH is considered the rate limiting enzyme for catecholamine synthesis, and serves to catalyze the hydroxylation of L-tyrosine. It maintains stores of catecholamines following secretion, and its activity is regulated by targeted site phosphorylation. Human TH is 528 amino acids (aa) in length. It contains an N-terminal ACT domain (aa 69-190) that binds small molecules and regulates enzyme activity, and a C-terminal enzymatic region (aa 196-493). There are three significant utilized phosphorylation sites. Two at Ser31 and Ser40 increase enzyme activity, while phosphorylation at Ser19 promotes subsequent Ser40 phosphorylation. TH functions as a 240 kDa noncovalent homotetramer. There are four potential splice variants. One shows a deletion of aa 31-34, a second shows a deletion of aa 31-61, while a third contains a Met substitution for aa 1-34. A fourth isoform variant shows a deletion of aa 35-61. Over aa 278-401, human TH shares 94% aa sequence identity with mouse TH, a molecule that most closely resembles the fourth human isoform variant described above.
  • Entrez Gene IDs:
    7054 (Human); 21823 (Mouse); 25085 (Rat)
  • Alternate Names:
    DYT14; DYT5b; EC 1.14.16; EC 1.14.16.2; TH; TYH dystonia 14; TYH; Tyrosine 3-hydroxylase; tyrosine 3-monooxygenase; Tyrosine Hydroxylase
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Average Rating: 4.5 (Based on 2 reviews)

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We have 2 reviews tested in 2 species: Human, Mouse.
We have 2 review tested in 1 application: Immunocytochemistry/Immunofluorescence.

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Images Ratings Applications Species Reviewed By Date Details
Immunocytochemistry/Immunofluorescence Human/Mouse Tyrosine Hydroxylase Antibody MAB7566
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Excellent
 ICC/IF Mouse Anonymous 10/27/2017
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Immunocytochemistry/Immunofluorescence Human/Mouse Tyrosine Hydroxylase Antibody MAB7566
Immunocytochemistry/Immunofluorescence: Human/Mouse Tyrosine Hydroxylase Antibody [MAB7566]

Summary

ApplicationImmunocytochemistry/Immunofluorescence
Sample TestedSympathetic nerve,conjunctival sympathetic nerve
SpeciesMouse
Immunocytochemistry/Immunofluorescence Human/Mouse Tyrosine Hydroxylase Antibody MAB7566
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Very Good
 ICC/IF Human Anonymous 10/24/2016
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Image Details
Immunocytochemistry/Immunofluorescence Human/Mouse Tyrosine Hydroxylase Antibody MAB7566
Immunocytochemistry/Immunofluorescence: Human/Mouse Tyrosine Hydroxylase Antibody [MAB7566]

Summary

ApplicationImmunocytochemistry/Immunofluorescence
Sample TestediPS2 human induced pluripotent stem cells
SpeciesHuman

Other Experimental Details

Other Experimental DetailsTH positive neurons in red, co-stained with TUJ1 to mark neurons.

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