Recombinant
RabMAb

Recombinant Anti-Tyrosine Hydroxylase antibody [EP1533Y] (ab75875)

Overview

  • Product name

    Anti-Tyrosine Hydroxylase antibody [EP1533Y]
    See all Tyrosine Hydroxylase primary antibodies
  • Description

    Rabbit monoclonal [EP1533Y] to Tyrosine Hydroxylase
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, Flow Cyt, IHC-Pmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Pig
  • Immunogen

    within Human Tyrosine Hydroxylase aa 50-150 (phospho S70). The exact sequence is proprietary.
    Database link: P07101
    (Peptide available as ab218000)

  • Positive control

    • Human adrenal gland lysate. This antibody gave a positive result in IF in the following Formaldehyde fixed cell line: SKNSH. IHC-P: Human cerebral cortex FFPE tissue sections.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab75875 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
ICC/IF 1/100 - 1/250.
WB 1/500. Predicted molecular weight: 59 kDa.Can be blocked with Tyrosine Hydroxylase peptide (ab183303) or Human Tyrosine Hydroxylase peptide (ab218000).
Flow Cyt 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P 1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for IP.
  • 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

    • Immunohistochemical analysis of rat brain tissue section labeling Tyrosine Hydroxylase with ab75875 at 1/1000 dilution. The tissue was fixed with Formaldehyde. Antigen retrieval was heat mediated using citric acid. A biotin conjugated goat anti-rabbit polyclonal IgG at 1/250 was used as the secondary antibody.

      See Abreview

    • Anti-Tyrosine Hydroxylase antibody [EP1533Y] (ab75875) at 1/500 dilution + human adrenal gland lysate at 10 µg

      Secondary
      Goat anti-rabbit HRP at 1/1000 dilution

      Predicted band size: 59 kDa
      Observed band size: 59 kDa

    • ab75875 staining Tyrosine Hydroxylase in Pig Myocardiun tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/1000 in blocking buffer) for 2 hours at 21°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.

      See Abreview

    • IHC image of Tyrosine Hydroxylase staining in human cerebral cortex formalin fixed paraffin embedded tissue section, performed on a Leica Bond 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 ab75875, 1/200 dilution, 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 ab75875 stained SKNSH cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab75875 at 1/100 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Overlay histogram showing SH-SY5Y cells stained with ab75875 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab75875, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    • Immunohistochemical analysis of mouse brain tissue section labeling Tyrosine Hydroxylase with ab75875 at 1/800 dilution. The tissue was fixed with Formaldehyde. Antigen retrieval was heat mediated using citric acid. A biotin conjugated goat anti-rabbit polyclonal IgG at 1/250 was used as the secondary antibody.

      See Abreview

    References

    This product has been referenced in:

    • Zhang L  et al. Semaglutide is Neuroprotective and Reduces a-Synuclein Levels in the Chronic MPTP Mouse Model of Parkinson's Disease. J Parkinsons Dis 9:157-171 (2019). Read more (PubMed: 30741689) »
    • Reeves FA  et al. Prostatic nerve subtypes independently predict biochemical recurrence in prostate cancer. J Clin Neurosci 63:213-219 (2019). Read more (PubMed: 30772200) »
    See all 15 Publications for this product

    Customer reviews and Q&As

    1-8 of 8 Abreviews or Q&A

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Nothobranchius furzeri Tissue sections (Brain)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate buffer pH 6.00
    Permeabilization
    Yes - Triton X-100 0.3% in PBS
    Specification
    Brain
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: RT°C
    Fixative
    Paraformaldehyde

    Miss. Adele Leggieri

    Verified customer

    Submitted Aug 10 2018

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (skeletal muscle)
    Permeabilization
    No
    Specification
    skeletal muscle
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Jul 20 2018

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: CC1
    Sample
    Human Tissue sections (Tonsil)
    Specification
    Tonsil
    Permeabilization
    No
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Jan 26 2015

    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 (Myocardium)
    Specification
    Myocardium
    Permeabilization
    No
    Fixative
    Formaldehyde

    Mr. Carl Hobbs

    Verified customer

    Submitted Nov 04 2014

    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
    Mouse Tissue sections (Brain)
    Specification
    Brain
    Permeabilization
    No
    Fixative
    Formaldehyde

    Mr. Carl Hobbs

    Verified customer

    Submitted Nov 01 2013

    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
    Rat Tissue sections (Brain)
    Specification
    Brain
    Permeabilization
    No
    Fixative
    Formaldehyde

    Mr. Carl Hobbs

    Verified customer

    Submitted Nov 01 2013

    Answer

    ab75875 and ab52493 are similar but not the same, they are different clones. They are both rabbit monoclonal antibodies. ab75875 was raised against a synthetic peptide corresponding to residues surrounding serine 70 of human Tyrosine Hydroxylase. ab52493 was raised against a synthetic peptide corresponding to residues near the N-terminus of human Tyrosinase.
    As shown on the product datasheet (https://www.abcam.com/ab75875), ab75875 has been tested in the following applications: Western Blot, ICC, flow cytometry, and in the following species: mouse, rat and human.

    If your customer wants to use this antibody in a species or application not listed on the datasheet, they may be eligible for a testing discount.

    Read More

    Question
    Answer

    Thank you for contacting Abcam regarding ab75875.


    Unfortunately, an ICC image for this antibody is not currently available. Therefore, I can offer a discount off a future purchase if you buy ab75875 now, use it in ICC and submit an image of your results to us in the form of an Abreview. The discount would be to the value of 1 free primary antibody.


    It doesn't matter whether the result is positive or negative, we would just really like to receive your feedback. Of course, if the results are negative you will be covered by our Abpromise guarantee and eligible for a refund or replacement.

    If you are interested in this offer, please follow these steps:

    1. Reply to this e-mail to let me know that you would like to proceed and send an image of ab75875 in ICC. I will then send a discount code. This code must be issued before purchasing ab75875 so please wait for my reply before ordering.

    2. Purchase ab75875 either by phone, fax, or online (www.abcam.com).

    3. Use it in ICC.

    4. Send us an image of the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

    5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue.

    We are always pleased to obtain feedback about our products and any information is greatly appreciated!

    Please let me know if you have any questions about this offer and I would be happy to help you further.

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

    Read More

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