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MAB5278 Anti-Tryptophan Hydroxylase/Tyrosine Hydroxylase/Phenylalanine Hydroxylase Antibody, clone PH8

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MAB5278
500 µg  
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      Tableau de caractéristiques principal

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      HIHC, IP, WBMPurifiedMonoclonal Antibody
      Description
      Catalogue NumberMAB5278
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      DescriptionAnti-Tryptophan Hydroxylase/Tyrosine Hydroxylase/Phenylalanine Hydroxylase Antibody, clone PH8
      Background InformationPH8 Antibody is a murine monoclonal antibody that binds a common epitope of Tryptophan hydroxylase (TRH), Tyrosine hydroxylase (TYH) and Phenylalanine hydroxylase (PAH).1 In the literature this monoclonal antibody is cited as PH8. Tyrosine hydroxylase (TYH) is the enzyme which converts tyrosine to dihydroxyphenylalanine (L-dopa), a precursor of the catecholamine neurotransmitters dopamine, noradrenaline and adrenaline. Tryptophan hydroxylase (TRH) is the enzyme that converts 5-hydroxytryptophan to serotonin. In fresh tissue the PH8 Antibody binds to TYH and TRH so is a marker for TYH-containing catecholaminergic neurons,2 and serotonergic neurons.3,4,5 Tryptophan hydroxylase (TRH) can be used as a marker for serotonin as it converts 5-hydroxy-tryptophan to serotonin. Serotonin is rapidly metabolised and is unable to be detected by anti-serotonin antibodies in post mortem tissue. In human tissue that has been formalin-fixed, due to a change in the antigenic determinant of tyrosine hydroxylase (TYH), PH8 Antibody will bind only tryptophan hydroxylase (TRH).3,4,5 The PH8 Antibody can therefore be used to specifically identify serotonergic neurons in fixed human tissue. Phenylalanine hydroxylase (PAH) can be detected in hepatic tissue sections using the PH8 Antibody.
      Product Information
      FormatPurified
      PresentationThe immunoglobulin fraction has been purified by Protein G chromatography. 500μg liquid protein at 2mg/mL in phosphate buffered saline (PBS). The product is supplied sterile-filtered through a 0.22 micron filter
      Quality LevelMQ100
      Applications
      ApplicationThis Anti-Tryptophan Hydroxylase/Tyrosine Hydroxylase/Phenylalanine Hydroxylase Antibody, clone PH8 is validated for use in IHC, IP, WB for the detection of Tryptophan Hydroxylase/Tyrosine Hydroxylase/Phenylalanine Hydroxylase.
      Key Applications
      • Immunohistochemistry
      • Immunoprecipitation
      • Western Blotting
      Application NotesThe PH8 Antibody can be used to identify dopaminergic and serotonergic neurons by immuno-histochemistry and for Western blot analysis, immunoprecipitation and immuno-histochemistry of TYH, PAH and TRH.

      PROTOCOLS

      Immunohistochemistry

      The PH8 Antibody can be used for the immunohistochemical detection of dopaminergic and serotonergic neurons in human and rat brain stem tissue.

      1. Tissue should be formalin fixed and stored in formalin prior to use for a minimum of five days.

      2. Cryoprotect tissue using 30% sucrose in 0.1M Tris pH 7.4 buffer for 24-72 hours.

      3. Cut using a sledge microtome to 50 μm thickness.

      4. Wash the tissue samples using Tris buffer prior to commencing the staining procedure.

      5. Treat the tissue samples for 3 x 15 minutes in 50% alcohol.

      6. Treat the tissue samples for 20 minutes in 50% alcohol and 3% H2O2.

      7. Treat tissue for 20 minutes with 10% normal horse serum in Tris buffer. This acts to block endogeneous H2O2 staining.

      8. Dilute the PH8 Antibody in Tris buffer, add to the tissue samples and incubate for 1-3 days. Recommended dilutions are 1:2,000-1:10,000. At high antibody concentration all monoaminergic neurons are stained with no distinction between serotonegic and catecholominergic cells. By diluting the antibody concentrate, cell types are distinguishable due to variation in staining intensity. At lower concentrations (1:5,000-1:10,000 dilution) only serotonergic cells will stain.

      9. Wash tissue (3 x 15 minutes), then add a biotinylated anti-mouse secondary antibody and incubate on an orbital shaker at room temperature (RT) for 1 hour.

      10. Wash tissue (3 x 15 minutes) and incubate on an orbital shaker at RT for 1 hour with the tertiary complex (ELITE KIT, Vector, USA).

      11. Wash tissue (3 x 15 minutes) and incubate on an orbital shaker at RT for 10 minutes with Tris buffered diamino-benzidine substrate. Add 0.1% H2O2 and incubate on an orbital shaker at RT for a further 5 minutes.

      12. Mount tissue onto gelatinised slides and allow to dry prior to microscopy.
      Biological Information
      ClonePH8
      HostMouse
      IsotypeIgG1
      Species Reactivity
      • Human
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryThis gene encodes a member of the pterin-dependent aromatic acid hydroxylase family. The encoded protein catalyzes the first and rate limiting step in the biosynthesis of serotonin, an important hormone and neurotransmitter. The human genome contains two related tryptophan hydroxylases, one on chromosome 11p15-p14 and one on chromosome 12q21. This gene is expressed predominantly in the gut, pineal gland, spleen, and thymus.
      Gene Symbol
      • TPH1
      • TPRH
      • MGC119994
      • TPH
      • TRPH
      • EC 1.14.16.4
      UniProt Number
      UniProt SummaryFUNCTION: 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 difficulty in walking and, in some cases, spasticity. Some cases present with parkinsonian symptoms in infancy and are referred to as autosomal recessive infantile parkinsonism.
      SIMILARITY: SwissProt: P07101 ## Belongs to the biopterin-dependent aromatic amino acid hydroxylase family.
      Product Usage Statements
      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.
      Storage and Shipping Information
      Storage ConditionsThe PH8 Antibody is shipped liquid at ambient temperature. Liquid material is stable for at up to 6 months when stored at 2-8°C.
      Packaging Information
      Material Size500 µg
      Global Trade Item Number
      Référence GTIN
      MAB5278 04053252464171