Overview

  • Product name
    Anti-Dopamine beta Hydroxylase antibody
    See all Dopamine beta Hydroxylase primary antibodies
  • Description
    Sheep polyclonal to Dopamine beta Hydroxylase
  • Host species
    Sheep
  • Tested applications
    Suitable for: ICC, WB, ICC/IF, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
  • Immunogen

    Synthetic peptide corresponding to Rat Dopamine beta Hydroxylase aa 43-62.
    Sequence:

    SEPPESPFPYHIPLDPEGTL

  • General notes
    Detects human, bovine, rat and mouse DBH. It is not effective for immunohistochemistry in human brain

Properties

Applications

Our Abpromise guarantee covers the use of ab19353 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 1/2500.
WB 1/5000. Detects a band of approximately 72 kDa.
ICC/IF 1/5000.
IHC-P 1/2000.
IHC-Fr Use at an assay dependent concentration.

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

  • Immunofluorescent staining of DBH in rat neuron cells using ab19353.

  • Immunofluorescent stain of DBH in rat brain tissue using ab19353.

References

This product has been referenced in:
  • Mahata SK  et al. Effect of heart failure on catecholamine granule morphology and storage in chromaffin cells. J Endocrinol 230:309-23 (2016). Read more (PubMed: 27402067) »
  • Talavera-Adame D  et al. Enhancement of embryonic stem cell differentiation promoted by avian chorioallantoic membranes. Tissue Eng Part A 15:3193-200 (2009). Read more (PubMed: 19364272) »
See all 2 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (Mouse brain and spinal cord)
Specification
Mouse brain and spinal cord

Abcam user community

Verified customer

Submitted Feb 22 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (Brain: Coronal sections of striatum and cortex)
Specification
Brain: Coronal sections of striatum and cortex

Abcam user community

Verified customer

Submitted Jan 28 2016

Question
Answer

DISCOUNT CODE: xxx
Expiration date: xxx
Value: xxx

I am very pleased to hear you would like to accept our offer and test ab19353 in IHC-Fr. This code will give you xxx off your next order before the expiration date. To redeem this offer, please submit an Abreview for IHC-Fr and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/abtrial.

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Question
Answer

Merci beaucoup de votre réponse et de vous avoir pris le temps de remplir le questionnaire.

Effectivement le point qui saute à l'œilen regardant leprotocole, est la méthode d'extraction de la protéine et préparation du lysat. J'ai aussi discuté ce point avec une collègue. Je vous suggère donc d'utiliser un tampon avec des détergents pour pouvoir extraire effectivement les protéines et pour ne pas devoir avoir une étape aussi longe de sonication et d'homogénéisation.

Je vous recommande donc fortement de préparer un lysat avec un tampon NP-40 pour s'assurer de l'extraction des protéines du cerveau?
Voici la composition du tampon et protocole que je peux recommander:

Nonidet-P40 (NP40) buffer

150 mM NaCl
1.0% NP-40 (possible to substitute with 0.1% Triton X-100)
50 mM Tris-HCl pH 8.0
Protease Inhibitors

Preparation of lysate from tissues

Dissect the tissue of interest with clean tools, on ice preferably, and as quickly as possible to prevent degradation by proteases.
Place the tissue in round-bottom microcentrifuge tubes or Eppendorf tubes and immerse in liquid nitrogen to “snap freeze”. Store samples at -80°C for later use or keep on ice for immediate homogenization. For a ˜5 mg piece of tissue, add ˜300 μl lysis buffer rapidly to the tube, homogenize with an electric homogenizer, rinse the blade twice with another 2 x 300 μl lysis buffer, then maintain constant agitation (for example, on an orbital shaker) for 2 hours at 4°C.
Centrifuge for 20 minutes at 16,000 x g at 4°C in a microcentrifuge for 20 minutes. Gently remove the tubes from the centrifuge and place on ice. Transfer the supernatant to a fresh tube kept on ice. Discard the pellet.



Pour plus d'information, veuillez ne pas hésiter à nous recontacter ou aussi de consulter notre protocole en ligne: https://www.abcam.com/index.html?pageconfig=resource&rid=13045

J'espère que cette suggestion va résoudre le problème. N'hésitez pas à me recontacter si vous souhaitez discuter un de ces points en plus de détails. Veuillez aussi me tenir au courant de vos résultats. Je vous remercie de votre collaboration.

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Question
Answer

Je suis désolée d'apprendre que ab19353 ne vous donne pas de bons résultats en Western blot.

Veuillez trouver ci-jointlequestionnaire discuté au téléphone qui nous permet de regrouper le plus d'informations possible sur le protocole que vous avez utilisé avec cet anticorps.
Celui-ci nous permettra de vous fournir la meilleure assistance technique.

Si nous concluons que notre produit est fautif et ne fonctionne pas comme explicité par notre fiche technique, nous serons ravis de mettre en place l'envoi d'un tube de remplacement gratuit ou d'un remboursement (sous réserve d'une commande ultérieure de 180 jours).

Je vous remercie par avance de nous envoyer votre protocole.

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Answer

Thank you for sending your protocol information. We haven't tested this antibody with perfusion-fixed or free floating tissue, but we generally recommend using an antigen retrieval method with formaldehyde-fixed tissue, as formaldehyde crosslinks proteins and the antibody binding sites can be masked. It is more difficult to perform heat-mediated retrieval with free-floating tissue, but you can try an enzymatic treatment with proteinase K or trypsin. We have some antigen retrieval protocols at the following link, just scroll down to the bottom of the page for enzymatic protocols- https://www.abcam.com/index.html?pageconfig=resource&rid=11488#3 I also recommend testing the secondary antibody with a different sheep primary antibody, to ensure that it is functional. I hope this information is helpful, but please let me know if you have further questions or concerns.

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