Overview

  • Product name
    Anti-Tyrosine Hydroxylase antibody
    See all Tyrosine Hydroxylase primary antibodies
  • Description
    Rabbit polyclonal to Tyrosine Hydroxylase
  • Host species
    Rabbit
  • Specificity
    Immunohistochemical distribution throughout the brain shows that staining is restricted to these neurons that are known to contain the enzyme.
  • Tested applications
    Suitable for: IHC-FoFr, ELISA, WB, ICC/IF, IHC (PFA fixed), IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Guinea pig, Human
    Predicted to work with: Chicken, Chimpanzee, Macaque monkey
  • Immunogen

    Synthetic peptide corresponding to Rat Tyrosine Hydroxylase aa 32-47 conjugated to Keyhole Limpet Haemocyanin (KLH) (Glutaraldehyde).
    Sequence:

    PRFIGRRQSLIEDARK


    Database link: P04177

  • Positive control
    • Mouse or rat brain containing dopaminergic neurons. Ventral mesencephalic neurons (mouse embryonic day 14).
  • General notes

     

    This antibody has proven useful in staining catecholaminergic neurons. It is a high quality reagent that stains these neurons intensely, including dendritic processes and fine nerve terminals.

Properties

Applications

Our Abpromise guarantee covers the use of ab6211 in the following tested applications.

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

Application Abreviews Notes
IHC-FoFr 1/500. PubMed: 19630993
ELISA Use at an assay dependent concentration.
WB 1/100 - 1/500. Predicted molecular weight: 42,53,84 kDa.
ICC/IF 1/1000.
IHC (PFA fixed) 1/1000.
IHC-Fr Use at an assay dependent concentration.

Target

  • Function
    Plays an important role in the physiology of adrenergic neurons.
  • Tissue specificity
    Mainly expressed in the brain and adrenal glands.
  • Pathway
    Catecholamine biosynthesis; dopamine biosynthesis; dopamine from L-tyrosine: step 1/2.
  • Involvement in disease
    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.
  • Sequence similarities
    Belongs to the biopterin-dependent aromatic amino acid hydroxylase family.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

Images

  • Tyrosine hydroxylase antibody (ab6211) staining rat mesencephalic serotoninergic neurons. Pictures taken using objectives X20 (A) or X20 (B). Note that the staining is observed in the cell body and the processes of these neurons. ab6211 was used at 1/1000 incubated for 2 days at room temperature. Secondary antibody was anti-rabbit Alexa Fluor 488 used at 1/1000 for 2 hours at room temperature. Rat brain tissue (ab29475) was perfusion fixed (paraformaldehyde 4% ) followed by post-fixation overnight in the same fixative, cryoprotected in 20% sucrose and then frozen in OCT. 30µm coronal sections of brain were cutting by cyrostat and immunohistochemistry was performed as the 'free floating' technique.

  • ab6211 staining tyrosine hydroxylase in PC-12 cells treated with orexin A (ab120212), by ICC/IF. Decrease in expression of tyrosine hydroxylase correlates with increased concentration of orexin A, as described in literature.
    The cells were incubated at 37°C for 1h in media containing different concentrations of ab120212 (orexin A) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum (ab7481), 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab6211 (1/1000 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

  • Anti-Tyrosine Hydroxylase antibody (ab6211) at 1/100 dilution + PC12 cell lysates at 6 µg

    Performed under reducing conditions.

    Predicted band size: 42,53,84 kDa



    Three bands are detected at ≈ 42 kDa, 53 kDa and 84 kDa.

  • ab6211 Rabbit polyclonal to Tyrosine Hydroxylase (1/1000) on cryopreserved ventral mesencephalic neurons dissected at mouse embryonic day 14. Neurons were thawed, plated and grown for 14 days, before fixation and overnight staining with ab6211.
  • Immunohistochemical detection of dopaminergic neurons in the rat zona incerta in a formalin-fixed floated cryostat section. Tyrosine hydroxylase immunoreactivity was visualized with ab6211 (1:100,000), using the biotinylated secondary antibody-ABC method and nickel-diaminobenzidine chromogen. Photo courtesy of Dr. Erik Hrabovszky, Hungarian Academy of Sciences, Budapest, Hungary.

References

This product has been referenced in:
  • Zhang X  et al. Autonomic reinnervation and functional regeneration in autologous transplanted submandibular glands in patients with severe keratoconjunctivitis sicca. Int J Oral Sci 10:14 (2018). Read more (PubMed: 29695713) »
  • Han X  et al. Plin4-Dependent Lipid Droplets Hamper Neuronal Mitophagy in the MPTP/p-Induced Mouse Model of Parkinson's Disease. Front Neurosci 12:397 (2018). Read more (PubMed: 29967574) »
See all 24 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Application
Immunohistochemistry (Resin sections)
Sample
Mouse Tissue sections (Retina)
Specification
Retina

Abcam user community

Verified customer

Submitted Jun 27 2016

Answer

Thank you for contacting Abcam.

I apologize for the delay, our laboratory was looking into the availability of the blocking peptide for ab6211.

We do not have the blocking peptide for ab6211 available separately. However, the peptide sequence is published: PRFIGRRQSLIEDARK, should you wish to pursue your own means of synthesizing the peptide. A 80-85% purity is generally good enough for blocking studies. Below is a link with a list of companies that provide peptide synthesis services:
http://www.peptideresource.com/custom-peptide.html

ab6211 recognizes amino acids 32-47 of Rat Tyrosine Hydroxylase, and is also reactive with mouse, guinea pig, and human tyrosine hydroxylase with predicted reactivity in chicken, chimpanzee, and macaque monkey. Thus, it should label total tyrosine hydroxylase through recognition of amino acids 32-47.

To our knowledge, ab6211 has not been tested in western blot. However by participating in our AbTrial program you can now use our products in an untested application or species without financial risk.

Simply follow these easy steps below to apply for our AbTrial Program:

1. Reply to this email, letting us know you are interested in testing this product.

2. Our scientists will email you an inactive personal discount code for the value of the product.

3. Purchase and test the product at the regular price.

4. Submit your results, including your discount code in the additional notes section of your Abreview.

5. Once the Abreview is submitted, the discount code will become active.

6. Apply your discount code on your next order to receive that value off.

The Terms and Conditions of this offer can be found at: https://www.abcam.com/abtrial

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Brain-SNpc)
Specification
Brain-SNpc
Fixative
Formaldehyde
Permeabilization
Yes - 0.3% TritonX in 0.1% PBS
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Apr 13 2012

Answer

Thank you for confirming this information and for your help and cooperation with this case. As requested, I have asked our accounting department to issue you with a credit note. This can then be redeemed against the invoice of a future order. As usual if you have any further questions regarding this credit note, please contact the accounts department by email at creditcontrol@abcam.com. The reference number, or credit ID, is ******. Please refer to this number in any correspondence with the accounting department. I would like to wish the customer good luck with their research. The technical team is always at your service, should you require further expert advice.  

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Question

LOT NUMBER 1077964 DESCRIPTION OF THE PROBLEM No staining SAMPLE Species: mouse What’s cell line or tissue: brain tissue PRIMARY ANTIBODY Species: rabbiy Reacts against: Tyrosine hydroxylase At what dilution(s) have you tested this antibody: 1:200/1:500/1:1000 Diluent buffer: 0.3% Triton X-100 Incubation time: overnight Incubation temperature: 4 degree What washing steps were done (which buffer, number of washes): PBS wash 3 time, each time 5 min ANTIBODY STORAGE CONDITIONS -20 FIXATION OF SAMPLE Yes or No : yes If yes, Fixative agent and concentration: paraformaldehyde Fixation time: perfusion and post-fix overnight Fixation temperature : 4 degree ANTIGEN RETRIEVAL no PERMEABILIZATION STEP Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers? yes Permeabilizing agent and concentration: PBS-0.1% Triton for 15 min at RT BLOCKING CONDITIONS Concentration: 10% donkey serum/PBS-0.05% Tween Blocking time : 1hr Blocking temperature: RT SECONDARY ANTIBODY Species: Alexa 594 donkey anti-rabbit Reacts against which species: At what dilution(s) have you tested this antibody: 1:200/1:500 Diluent buffer: 0.3% Triton X-100 Incubation time: overnight What washing steps were done (which buffer, number of washes): PBS wash 3 time, each time 5 min ˙Fluorochrome or enzyme conjugate (eg: FITC, HRP, AP, biotin…etc): Alexa 594 donkey anti-rabbit Do you know whether the problems you are experiencing come from the secondary? NO HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 7 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? NO

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Answer

Thank you for filling in the questionnaire and for reporting this problem to us. We will use the details you sent about your protocol in order to investigate the source of the problem. I have looked through your protocol, which looks fine to me. I appreciate the time that has been spent on these experiments and would be pleased to arrange a credit note or a replacement. Unfortunately the only lot of ab6211 we currently have in stock is the same your customer received. Therefore I can offer a replacement with one of the following alternative products, tested in Mouse and IHC-FoFR : ab41528, www.abcam.com/ab41528 ab51191, www.abcam.com/ab51191 ab112, www.abcam.com/ab112 ab113, www.abcam.com/ab113 I look forward to hearing from you with details of how you would like to proceed. Could you also please inform us that you added glycerol to ab6211 before storing at -20°C as recommended on the datasheet?

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Answer

Thank you for your enquiry. Since you are seeing some specific staining in addition to this ubiquitous background staining, I am thinking this may be non-specific staining due to using too high a concentration of primary antibody. The recommendation on the datasheet is a starting point only, and I would recommend diluting this antibody out further, perhaps trying serial dilutions 1:2000 and 1:4000. In addition, I didn't see from your enquiry how you were incubating with the antibody. I would recommend an overnight incubation at 4C, as this will give the most specific signal. If you are still having problems with this antibody after following these suggestions, please let us know. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for your enquiry. Regarding "using protein in the phenol-ethanol supernatant for ELISA", would you first precipitate the protein? In either case, I'm not sure if the protein would be recognized by the antibodies since we have not tested this ourselves. We invite you to use our Abreview system. The Abreview system allows researchers to submit reviews of our products in exchange for Abpoints. Abpoints can be redeemed for rewards such as Amazon.com gift certificates or discounts on future antibody purchases. Should you decide to go ahead and purchase this product, please let us know how you get on by submitting an Abreview right from the online datasheet. I hope this information helps and please let me know how the ELISAs work.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Rat mesencephalic brain)
Specification
Rat mesencephalic brain
Fixative
Paraformaldehyde

Dr. Sophie Pezet

Verified customer

Submitted Apr 12 2006

Answer

Thank you for your enquiry. All the information we have on species cross reactivity is specified on the datasheet, these are updated as soona s any new information is brought to our attention. As far as we are aware, cross reactivity with human has not yet been tested for use with ab6211. Should you decide to go ahead and purchase this product, please let us know how you get on and in return we will forward a reward of your choice, typically an Amazon gift voucher. Please let me know if you require any further assistance.

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Question
Answer

There are no preservatives present in the whole serum. The whole antiserum was just sterilized by pushing it through a 0.2 um amicon filter.

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1-10 of 11 Abreviews or Q&A

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