Recombinant
RabMAb

Recombinant Anti-Dopamine beta Hydroxylase antibody [EPR20385] (ab209487)

Overview

  • Product name

    Anti-Dopamine beta Hydroxylase antibody [EPR20385]
    See all Dopamine beta Hydroxylase primary antibodies
  • Description

    Rabbit monoclonal [EPR20385] to Dopamine beta Hydroxylase
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human Dopamine beta Hydroxylase aa 50-350. The exact sequence is proprietary.
    Database link: P09172

  • Positive control

    • WB: SH-SY5Y whole cell lysate; Mouse and rat adrenal gland lysates. IHC-P: Human adrenal gland and adrenal pheochromocytoma tissues; Mouse and rat adrenal gland tissues. IP: SH-SY5Y whole cell lysate.
  • 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab209487 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
WB 1/1000. Detects a band of approximately 75 kDa (predicted molecular weight: 69 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tris/EDTA Buffer, pH9 (ab93684)

IP 1/40.

Target

  • Function

    Conversion of dopamine to noradrenaline.
  • Pathway

    Catecholamine biosynthesis; (R)-noradrenaline biosynthesis; (R)-noradrenaline from dopamine: step 1/1.
  • Involvement in disease

    Defects in DBH are the cause of dopamine beta-hydroxylase deficiency (DBH deficiency) [MIM:223360]; also known as norepinephrine deficiency or noradrenaline deficiency. This disorder is characterized by profound deficits in autonomic and cardiovascular function, but apparently only subtle signs, if any, of central nervous system dysfunction.
  • Sequence similarities

    Belongs to the copper type II ascorbate-dependent monooxygenase family.
    Contains 1 DOMON domain.
  • Cellular localization

    Cytoplasmic vesicle > secretory vesicle lumen. Cytoplasmic vesicle > secretory vesicle > chromaffin granule lumen and Cytoplasmic vesicle > secretory vesicle membrane. Cytoplasmic vesicle > secretory vesicle > chromaffin granule membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • dbh antibody
    • DBM antibody
    • Dopamine beta hydroxylase antibody
    • Dopamine beta monooxygenase antibody
    • Dopamine beta-hydroxylase (dopamine beta-monooxygenase) antibody
    • Dopamine beta-monooxygenase antibody
    • DOPO_HUMAN antibody
    • OTTHUMP00000022501 antibody
    • Soluble dopamine beta-hydroxylase antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded human adrenal gland tissue labeling Dopamine beta Hydroxylase with ab209487 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Strong cytoplasmic staining on chromaffin cells of human adrenal gland medulla, and negative on adrenal cortex (PMID: 17535872, PMID: 17699566).

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • All lanes : Anti-Dopamine beta Hydroxylase antibody [EPR20385] (ab209487) at 1/1000 dilution

    Lane 1 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 3 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 4 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 69 kDa
    Observed band size: 75 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Negative controls: HeLa (PMID: 7688735; PMID: 9763470, PMID:15585414, PMID: 10037744), C6 (PMID: 9763470, PMID:7798950, PMID: 10037744), HepG2 (PMID:7798950, PMID:11943777, PMID:15585414).

  • Dopamine beta Hydroxylase was immunoprecipitated from 0.35 mg of SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate with ab209487 at 1/40 dilution.

    Western blot was performed from the immunoprecipitate using ab209487 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: SH-SY5Y whole cell lysate, 10 µg (Input).

    Lane 2: ab209487 IP in SH-SY5Y whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab209487 in SH-SY5Y whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Immunohistochemical analysis of paraffin-embedded human adrenal pheochromocytoma tissue labeling Dopamine beta Hydroxylase with ab209487 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic staining on tumor cells of human adrenal pheochromocytoma is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • All lanes : Anti-Dopamine beta Hydroxylase antibody [EPR20385] (ab209487) at 1/1000 dilution

    Lane 1 : Mouse adrenal gland lysate
    Lane 2 : Rat adrenal gland lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 69 kDa
    Observed band size: 75 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 3 minutes; Lane 2: 1 second.

  • Immunohistochemical analysis of paraffin-embedded mouse adrenal gland tissue labeling Dopamine beta Hydroxylase with ab209487 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic staining on chromaffin cells of mouse adrenal gland medulla, and negative on adrenal cortex (PMID: 17535872, PMID: 176995660).

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded rat adrenal gland tissue labeling Dopamine beta Hydroxylase with ab209487 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic staining on chromaffin cells of rat adrenal gland medulla, and negative on adrenal cortex (PMID: 17535872, PMID: 176995660).

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

References

This product has been referenced in:

  • Li Y  et al. Investigation of Behavioral Dysfunctions Induced by Monoamine Depletions in a Mouse Model of Parkinson's Disease. Front Cell Neurosci 12:241 (2018). Read more (PubMed: 30135645) »
See 1 Publication for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Colon)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Colon
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Nov 21 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Adrenal gland)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Adrenal gland
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Nov 21 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Nov 21 2018

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
None
Permeabilization
Yes - 0.3% Triton X-100
Specification
Brain
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5 · Temperature: 24°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 10 2018

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