Millipore Sigma Vibrant Logo
Attention: We have moved. Merck Millipore products are no longer available for purchase on MerckMillipore.com.Learn More

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

View Products on Sigmaaldrich.com
MAB5278
500 µg  
Purchase on Sigma-Aldrich

Special Offers

Overview

Replacement Information

Key Spec Table

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.
References
Product Information
FormatPurified
HS Code3002 15 90
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.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
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
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
MAB5278 04053252464171

Documentation

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

Title

Safety Data Sheet (SDS) 

Anti-Tryptophan Hydroxylase/Tyrosine Hydroxylase/Phenylalanine Hydroxylase Antibody, clone PH8 Certificates of Analysis

TitleLot Number
PH 8 Antibody-anti human Tryptophan Hydroxylase, Tyrosine Hydroxylase and Phenylalanine Hydroxylase - 3213926 3213926
PH 8 Antibody-anti human Tryptophan Hydroxylase, Tyrosine Hydroxylase and Phenylalanine Hydroxylase - 3684759 3684759

References

Reference overviewApplicationSpeciesPub Med ID
5-HT neurons of the area postrema become c-Fos-activated after increases in plasma sodium levels and transmit interoceptive information to the nucleus accumbens.
Miller, RL; Loewy, AD
American journal of physiology. Regulatory, integrative and comparative physiology  306  R663-73  2014

Show Abstract
24598462 24598462
A new model for allosteric regulation of phenylalanine hydroxylase: implications for disease and therapeutics.
Jaffe, EK; Stith, L; Lawrence, SH; Andrake, M; Dunbrack, RL
Archives of biochemistry and biophysics  530  73-82  2013

Show Abstract
Western Blotting23296088 23296088
ENaC-expressing neurons in the sensory circumventricular organs become c-Fos activated following systemic sodium changes.
Miller, RL; Wang, MH; Gray, PA; Salkoff, LB; Loewy, AD
American journal of physiology. Regulatory, integrative and comparative physiology  305  R1141-52  2013

Show Abstract
24049115 24049115
Brainstem deficiency of the 14-3-3 regulator of serotonin synthesis: a proteomics analysis in the sudden infant death syndrome.
Broadbelt, KG; Rivera, KD; Paterson, DS; Duncan, JR; Trachtenberg, FL; Paulo, JA; Stapels, MD; Borenstein, NS; Belliveau, RA; Haas, EA; Stanley, C; Krous, HF; Steen, H; Kinney, HC
Molecular & cellular proteomics : MCP  11  M111.009530  2012

Show Abstract
Human21976671 21976671
Axonal inclusions in spinocerebellar ataxia type 3.
Seidel, K; den Dunnen, WF; Schultz, C; Paulson, H; Frank, S; de Vos, RA; Brunt, ER; Deller, T; Kampinga, HH; Rüb, U
Acta neuropathologica  120  449-60  2010

Show Abstract
20635090 20635090
5-HT2A receptors are concentrated in regions of the human infant medulla involved in respiratory and autonomic control.
David S Paterson, Ryan Darnall, David S Paterson, Ryan Darnall, David S Paterson, Ryan Darnall, David S Paterson, Ryan Darnall
Autonomic neuroscience : basic clinical  147  48-55  2009

Show Abstract Full Text Article
19213611 19213611
The development of nicotinic receptors in the human medulla oblongata: inter-relationship with the serotonergic system.
Duncan, JR; Paterson, DS; Kinney, HC
Autonomic neuroscience : basic & clinical  144  61-75  2008

Show Abstract
18986852 18986852
Activation and stabilization of human tryptophan hydroxylase 2 by phosphorylation and 14-3-3 binding.
Ingeborg Winge,Jeffrey A McKinney,Ming Ying,Clive S D'Santos,Rune Kleppe,Per M Knappskog,Jan Haavik
The Biochemical journal  410  2008

Show Abstract
17973628 17973628
Neuronal expression of constitutive heat shock proteins: implications for neurodegenerative diseases.
Chen, S; Brown, IR
Cell stress & chaperones  12  51-8  2007

Show Abstract Full Text Article
17441507 17441507