Recombinant
RabMAb

Anti-Tyrosine Hydroxylase antibody [EP1533Y] (Alexa Fluor® 488) (ab192463)

Overview

  • Product name
    Anti-Tyrosine Hydroxylase antibody [EP1533Y] (Alexa Fluor® 488)
    See all Tyrosine Hydroxylase primary antibodies
  • Description
    Rabbit monoclonal [EP1533Y] to Tyrosine Hydroxylase (Alexa Fluor® 488)
  • Host species
    Rabbit
  • Conjugation
    Alexa Fluor® 488. Ex: 495nm, Em: 519nm
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, IHC-Frmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Tyrosine Hydroxylase aa 50-150 (phospho S70). The exact sequence is proprietary.
    Database link: P07101

  • Positive control
    • ICC/IF: SKNSH cells. Flow Cyt: SH-SY5Y cells. IHC-Fr: Rat frozen dentate gyrus tissue section
  • 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.

    Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

Properties

Applications

Our Abpromise guarantee covers the use of ab192463 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
Flow Cyt 1/500.

ab199091 - Rabbit monoclonal IgG (Alexa Fluor® 488), is suitable for use as an isotype control with this antibody.

ICC/IF 1/50.
IHC-Fr 1/50.

This product gave a positive signal in rat frozen dentate gyrus tissue fixed with 10% formaldehyde (10 minutes).

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

  • IHC image of Tyrosine Hydroxylase staining in a section of frozen normal rat dentate gyrus.

    The section was fixed using 10% formaldehyde in 1XPBS for 10 minutes. No antigen retrieval step was performed prior to staining. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab192463 at 1/50 (shown in green) and counterstained using ab195884, Rat monoclonal to Tubulin (Alexa Fluor® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount®.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

  • Overlay histogram showing SH-SY5Y cells stained with ab192463 (red line). The cells were fixed with 4% formaldehyde (10 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. 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 (ab192463, 1/500 dilution) for 30 minutes at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor® 488 (ab199091) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 minutes)/permeabilized with 0.1% PBS-Tween for 20 minutes used under the same conditions.

  • ab192463 staining Tyrosine Hydroxylase in SKNSH cells. The cells were fixed with 4% formaldehyde (10 minutes), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated with ab192463 at a working dilution of 1 in 50 (shown in green) and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 594, shown in red) at 2 µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

References

This product has been referenced in:
  • Kiedrowski L & Feinerman A Medium-retaining Petri dish insert to grow and image cultured cells. J Neurosci Methods 294:111-115 (2018). Read more (PubMed: 29132810) »
  • Shi J  et al. AM1241 alleviates MPTP-induced Parkinson's disease and promotes the regeneration of DA neurons in PD mice. Oncotarget 8:67837-67850 (2017). IHC . Read more (PubMed: 28978077) »
See all 2 Publications for this product

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (primary culture of ventral mesencephalon cells)
Permeabilization
Yes - 0.3% Triton X-100
Specification
primary culture of ventral mesencephalon cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde

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Verified customer

Submitted Aug 24 2015

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