Human/Mouse/Rat Tyrosine Hydroxylase Antibody

  
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human, mouse and rat Tyrosine Hydroxylase in Western blots. In direct ELISAs, less than 5% cross-reactivity with recombinant human TPH1 is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human Tyrosine Hydroxylase
    Ala278-Tyr401, predicted
    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
    1 µg/mL
    See below
  • Simple Western
    10 µg/mL
    See below
  • Immunocytochemistry
    5-15 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 PC‑12 rat adrenal pheochromocytoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Tyrosine Hydroxylase Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7566) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for Tyrosine Hydroxylase at approximately 56-60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry

Tyrosine Hydroxylase in D3 Mouse Embryonic Stem Cells. Tyrosine Hydroxylase was detected in immersion fixed D3 mouse embryonic stem cells using Sheep Anti-Human Tyrosine Hydroxylase Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7566) at 10 µg/mL for 3 hours at room temperature. Cells were differentiated into dopaminergic neurons using R&D Systems Human/Mouse Dopaminergic Neuron Differentiation Kit (Catalog # SC001B). Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to dopaminergic neurons. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Rat Tyrosine Hydroxylase by Simple WesternTM. Simple Western lane view shows lysates of PC‑12 rat adrenal pheochromocytoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Tyrosine Hydroxylase at approximately 59 kDa (as indicated) using 10 µg/mL of Sheep Anti-Human/Mouse/Rat Tyrosine Hydroxylase Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7566) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.

Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.2 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 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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