MAB5278
Anti-Tryptophan Hydroxylase/Tyrosine Hydroxylase/Phenylalanine Hydroxylase Antibody
CHEMICON®, mouse monoclonal, PH8
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About This Item
Kod UNSPSC:
12352203
eCl@ss:
32160702
NACRES:
NA.41
Polecane produkty
Nazwa produktu
Anti-Tryptophan Hydroxylase/Tyrosine Hydroxylase/Phenylalanine Hydroxylase Antibody, clone PH8, clone PH8, Chemicon®, from mouse
pochodzenie biologiczne
mouse
Poziom jakości
forma przeciwciała
purified immunoglobulin
rodzaj przeciwciała
primary antibodies
klon
PH8, monoclonal
reaktywność gatunkowa
human
producent / nazwa handlowa
Chemicon®
metody
immunohistochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable
izotyp
IgG1
numer dostępu NCBI
Warunki transportu
wet ice
docelowa modyfikacja potranslacyjna
unmodified
informacje o genach
human ... TPH1(7166)
Opis ogólny
PH8 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.
Zastosowanie
The 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.
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.
This 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.
Postać fizyczna
Format: Purified
The immunoglobulin fraction has been purified by Protein G chromatography. 500?g liquid protein at 2mg/mL in phosphate buffered saline (PBS). The product is supplied sterile-filtered through a 0.22 micron filter
Przechowywanie i stabilność
The PH8 Antibody is shipped liquid at ambient temperature. Liquid material is stable for at up to 6 months when stored at 2-8°C.
Informacje prawne
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
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Kod klasy składowania
12 - Non Combustible Liquids
Klasa zagrożenia wodnego (WGK)
WGK 2
Temperatura zapłonu (°F)
Not applicable
Temperatura zapłonu (°C)
Not applicable
Certyfikaty analizy (CoA)
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Dokumenty związane z niedawno zakupionymi produktami zostały zamieszczone w Bibliotece dokumentów.
Fiona M Bright et al.
Journal of neuropathology and experimental neurology, 76(10), 864-873 (2017-09-20)
Serotonin (5-hydroxytryptamine [5-HT]) neurons in the medulla oblongata project extensively to key autonomic and respiratory nuclei in the brainstem and spinal cord regulating critical homeostatic functions. Multiple abnormalities in markers of 5-HT function in the medulla in sudden infant death
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 null
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 null
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 et al.
Autonomic Neuroscience : Basic & Clinical null
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 null
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