Anti-Tyrosine Hydroxylase抗体- Neuronal Marker (ab41528)

概述

  • 产品名称Anti-Tyrosine Hydroxylase抗体- Neuronal Marker
    参阅全部 Tyrosine Hydroxylase 一抗
  • 描述
    兔多克隆抗体to Tyrosine Hydroxylase - Neuronal Marker
  • 经测试应用适用于: IHC-FoFr, WB, ICC/IF, IHC-Fr, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Pig
    预测可用于: Human
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 400 - 500 of Human Tyrosine Hydroxylase.

    (Peptide available as ab41527)

  • 阳性对照
    • ab41528 gave a positive result in the following tissue lysates: Mouse Substantia Nigra, Rat Substantia Nigra, Mouse Dopaminergic Nucleus and Rat Dopaminergic Nucleus. This antibody gave a positive result in IHC in the following FFPE tissue: Rat 6 week brain.

性能

相关产品

应用

Our Abpromise guarantee covers the use of ab41528 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
IHC-FoFr Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 59 kDa).Can be blocked with Human Tyrosine Hydroxylase peptide (ab41527).
ICC/IF Use a concentration of 1 µg/ml.
IHC-Fr 1/200 - 1/400.
IHC-P Use a concentration of 1 µg/ml.

靶标

  • 功能Plays an important role in the physiology of adrenergic neurons.
  • 组织特异性Mainly expressed in the brain and adrenal glands.
  • 通路Catecholamine biosynthesis; dopamine biosynthesis; dopamine from L-tyrosine: step 1/2.
  • 疾病相关Defects in TH are the cause of dystonia DOPA-responsive autosomal recessive (ARDRD) [MIM:605407]; also known as autosomal recessive Segawa syndrome. ARDRD is a form of DOPA-responsive dystonia presenting in infancy or early childhood. Dystonia is defined by the presence of sustained involuntary muscle contractions, often leading to abnormal postures. Some cases of ARDRD present with parkinsonian symptoms in infancy. Unlike all other forms of dystonia, it is an eminently treatable condition, due to a favorable response to L-DOPA.
    Note=May play a role in the pathogenesis of Parkinson disease (PD). A genome-wide copy number variation analysis has identified a 34 kilobase deletion over the TH gene in a PD patient but not in any controls.
  • 序列相似性Belongs to the biopterin-dependent aromatic amino acid hydroxylase family.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Dystonia 14 antibody
    • DYT14 antibody
    • DYT5b antibody
    • EC 1.14.16.2 antibody
    • OTTHUMP00000011225 antibody
    • OTTHUMP00000011226 antibody
    • ple antibody
    • Protein Pale antibody
    • TH antibody
    • The antibody
    • TY3H_HUMAN antibody
    • TYH antibody
    • Tyrosine 3 hydroxylase antibody
    • Tyrosine 3 monooxygenase antibody
    • Tyrosine 3-hydroxylase antibody
    • Tyrosine 3-monooxygenase antibody
    • Tyrosine hydroxylase antibody
    see all

Anti-Tyrosine Hydroxylase antibody - Neuronal Marker 图像

  • All lanes : Anti-Tyrosine Hydroxylase antibody - Neuronal Marker (ab41528) at 1 µg/ml

    Lane 1 : Mouse Substantia Nigra at 5 µg
    Lane 2 : Rat Substantia Nigra at 10 µg
    Lane 3 : Mouse Dopaminergic Nucleus at 10 µg
    Lane 4 : Rat Dopaminergic Nucleus at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 59 kDa
    Observed band size : 60 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 70 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 30 seconds
  • IHC image of Tyrosine Hydroxylase staining in Rat 6 week brain (coronal) formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab41528, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • ICC/IF image of ab41528 stained rat PC12 cells. The cells were methanol fixed (5 min), permabilised in PBS-T (20 min) and incubated with the antibody (ab41528, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

  • Minipigs were deeply anesthetized with a combination of midazolam and ketamine, prior to transcardial perfusion with phosphate buffered 4% paraformaldehyde (pH 7.4). After perfusion, the brains were removed with special care taken to preserve the optic chiasm and the median eminence. Blocks of tissue containing the hypothalami were dissected, postfixed in the same fixative for 1 day and subsequently cryoprotected in 30% sucrose for 3–4 days, prior to freezing. 10 series of 40-mm thick coronal (6 animals), sagittal (1 animal), and horizontal (1 animal) cryostat sections were collected. Coronal sections for immunohistochemistry were maintained at -18°C as free-floating sections in a cryoprotectant poly-ethylene glycol solution for up to four weeks.
    Immunohistochemistry was performed using the avidin-biotin method. Accordingly, free-floating sections were first rinsed in Tris-buffered saline (TBS; 0.05 M; pH 7.4) for 15 minutes. Incubations with avidin (0.1%) and biotin (0.01%) were each of 10 minute duration and each procedure were followed by a 2 minute TBS-rinse. The sections were preincubated with 1% Triton X- 100 and 0.2% milk in TBS for 30 minutes, prior to incubation with the primary antibody, ab41528, for 72 hours at 4°C at a 1/400 dilution. The sections were then washed three times 15 minutes in TBS and 1% Triton X-100 prior to incubation for 1 hour at room temperature with the secondary anti-rabbit biotinylated antibody at a 1/400 with TBS that contained 1% Triton X- 100 and 0.2% milk. After a brief rinse in TBS, the endogenous peroxidase activity was blocked with a solution of 10 ml hydrogen peroxide, 10 ml methanol, and 80 ml TBS, for 10 minutes. The sections were then rinsed in TBS and 1% Triton for three times 15 minutes and incubated for 1 hour at room temperature with avidin-peroxidas diluted 1/400 in TBS that contained 1% Triton X-100 and 0.2% milk. The sections were then rinsed three times for 15 minutes in TBS and 1% Triton, prior to avidin-peroxidase visualization. The latter step consisted of incubation for 10 minutes in 10ml water in which one diaminobenzidine tablet had been dissolved and to which 10 ml of 35% H2O2 had been added. Finally, the sections were dehydrated, mounted and coverslipped with Depex.

    PHA: posterior hypothalamic area.

    VMH: ventromedial hypothalamic nucleus.

    Arc: arcuate nucleus.

Anti-Tyrosine Hydroxylase antibody - Neuronal Marker (ab41528)参考文献

This product has been referenced in:
  • Yu J  et al. Effects of Simazine Exposure on Neuronal Development-Related Factors in MN9D Cells. Med Sci Monit 22:2831-8 (2016). WB ; Mouse . Read more (PubMed: 27513680) »
  • Ettrup KS  et al. The anatomy of the Göttingen minipig hypothalamus. J Chem Neuroanat 39:151-65 (2010). IHC-FoFr ; Pig . Read more (PubMed: 20043984) »

See all 2 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Sample Pig Tissue sections (Heart muscle)
Specification Heart muscle
Permeabilization No
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

提交于 Nov 04 2014

Thank you for contacting us.

This antibody is tested for specificity towards Pig protein so it is guaranteed. Although many vials of this product has been sold however we are aware of only one publication; Ettrup KS et al. The anatomy of the G...

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According to our records, ab41528 was proving difficult to use and we were in contact in order to help resolve the issue.

Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the d...

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Thank you very much for your call yesterday and for your question about this antibody.

The lab has not further characterized the˜70 kDa band that is seen in the Western blot image, but the lab scientists believe it to be non-specific. Though ...

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Thank you for contacting Abcam. Please find the protocol used for culturing the PC12 cells below: We grow the PC12 cells in Invitrogen Gibco RPMI media with 5% FBS, 2mM L-glutamine, 10% Horse Serum, and 2.5mL Penicillin/Streptomycin antibiotic. PC12 ...

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Thank you for contacting Abcam yesterday. I am still waiting to hear back about how the PC12 cells were cultured but I have found out that the cells were cultured on polylyseine coated coverslips. I will get back to you as soon as I get more inform...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Kidney and Adrenal gland)
Specification Kidney and Adrenal gland
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

提交于 Aug 02 2011

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Colon)
Specification Colon
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1%
Username

Mr. Carl Hobbs

Verified customer

提交于 Aug 02 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (brain sections)
Specification brain sections
Fixative Paraformaldehyde
Permeabilization No
Username

Dr. Sophie Pezet

Verified customer

提交于 Jul 09 2008

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"