Overview

  • Product name

  • Description

    Rabbit polyclonal to Dopamine
  • Host species

    Rabbit
  • Specificity

    Using a conjugate Dopamine-Glutaraldehyde-bovine serum albumin, antibody specificity was performed with an ELISA test by competition experiments with the following compounds:
    G-BSA-conjugateCross-reactivity ratio (a)
    Dopamine1
    Dopamine(b)1/2.5
    Noradrenaline1/159
    L-DOPA, Octopamine1/400
    Tyramine, Adrenalin, unconjugated Dopamine 1/>5,000
    (a): Dopamine-G-BSA concentration/unconjugated or conjugated catecholamine concentration at half displacement (b): Non-reduced conjugate
  • Tested applications

    Suitable for: ICC, ICC/IF, ELISAmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Chemical/ Small Molecule by a Glutaraldehyde linker.

  • General notes

    The antiserum was tested using the free floating PAP technique on rat dopaminergic areas. The anti-conjugated Dopamine antibodies gave good staining between 1/2000-1/5000 dilution in these areas.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Concentration information loading...
  • Purity

    Ammonium Sulphate Precipitation
  • Primary antibody notes

    The antiserum was tested using the free floating PAP technique on rat dopaminergic areas. The anti-conjugated Dopamine antibodies gave good staining between 1/2000-1/5000 dilution in these areas.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab6427 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 Use at an assay dependent concentration.
ICC/IF 1/1000 - 1/5000. Glutharaldehyde perfusion fixation (see recommended protocol).
ELISA 1/5000.

Target

  • Relevance

    Dopamine (C6H3OH)2-CH2CH2NH2)is a catecholamine neurotransmitter in the brain. Its chemical name is 4-(2-aminoethyl)benzene-1,2-diol. Dopamine is a hormone released by the hypothalamus. Its main function is to inhibit the release of prolactin from the anterior lobe of the pituitary. It can be used as a sympathomimetic drug, producing effects such as increased heart rate and blood pressure.
  • Cellular localization

    Secreted
  • Alternative names

    • 3 4 dihydroxyphenethylamine antibody
    • 3 hydroxytyramine antibody
    • DA antibody
    • Dopamine antibody
    • Oxytyramine antibody
    see all

Images

  • ab6427 at 1/5000 staining human stem cells (differentiated to neurons) by ICC. The cells were paraformaldehyde fixed and blocked with BSA before incubation with the antibody for 2 hours. A PE conjugated goatanti-rabbit antibody was used as the secondary.

    See Abreview

  • ab6427 staining dopamine in PC12 cells treated with ghrelin (rat) (ab120231), by ICC/IF. Decrease in dopamine expression correlates with increased concentration of ghrelin, as described in literature.
    The cells were incubated at 37°C for 10 minutes in media containing different concentrations of ab120231 (ghrelin) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab6427 (1/200 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.

References

This product has been referenced in:

  • Fink J  et al. Development of a Competition-Binding Assay to Determine Binding Affinity of Molecules to Neuromelanin via Fluorescence Spectroscopy. Biomolecules 9:N/A (2019). Read more (PubMed: 31072013) »
  • Taylor-Whiteley TR  et al. Recapitulating Parkinson's disease pathology in a three-dimensional human neural cell culture model. Dis Model Mech 12:N/A (2019). Read more (PubMed: 30926586) »
See all 6 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer

Thank you for contacting us.

ab6427 is not guaranteed for use in immunohistochemistry. However, ab1001 (https://www.abcam.com/ab1001) is guaranteed to work in immunohistochemistry and gave good results when fixed with formalin/PFA in paraffin-embedded sections.

Regarding your question about whether this antibody works on rat tissue, because the immunogen of ab1001 and ab6427 is a Chemical / Small Molecule, the species in which this antibody is used is irrelevant. Therefore both these antibodies should work on rat tissue.

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

Read More
Application
Immunocytochemistry
Sample
Human Cell (stem cells differentiated to neurons)
Specification
stem cells differentiated to neurons
Fixative
Paraformaldehyde
Blocking step
BSA as blocking agent for 45 minute(s) · Concentration: 2

Abcam user community

Verified customer

Submitted May 29 2007

Answer

We have additional information about capturing the DA as follows: After discussion with our research team, we think that the better way is to bind DA on your tissue via glutaraldehyde (0.1 to 1%). - Double bounds are reduced by adding of sodium borohydrure. - Tissue has to be wash for eliminate borohydrure excess. - You have also to make a competition scale between antibody and different concentration of the antigen DA-G-BSA. Had this medium on the tissue. - After revelation by a labelled anti-rabbit antibody, you finally have to estimate the competition rate between DA linked on tissue and soluble DA-G-BSA conjugate.

Read More

Answer

Thank you for your enquiry. For Immunohistological studies with dopamine antibodies, fixation with glutaraldehyde is required. Please find below a protocol. Perfusion protocol for adult male Sprague-Dawley (weight around 0.5 kg) 1- The animals can be deeply anaesthetized for example with uerethane (0.1-1.5g/kg, intraperitoneal). 2- Heparinized and perfused via the ascending aorta with 100 ml of cold physiologic saline (0.9% NaCl) and with the following fixative solution: a. 300 ml of cold 4% paraformaldehyde and 2% glutaraldehyde in 0.1M Phosphate buffer (PB), pH 7.2 ( in two minutes). b. 600 ml of cold 4% paraformaldehyde and 2% glutaraldehyde in 0.1M PB, pH 7.2 ( in ten minutes). c. Dissect out the brains and place in a solution of 4% paraformaldehyde in 0.1M PB, pH 7.2, at 4°C for twelve to sixteen hours. d. Before brain will be cut on a freezing microtome, we must include it in growing concentrations of sucrose (5 at 30 %). If you have any further questions then please do not hesitate to get back in touch with us.

Read More

Question
Answer

The pump that was used is a peristaltic pump with a flow of 150-300ml/minute. GILSON supply this pump type.

Read More

Answer

This antibody is known to work with 3-5% glutaraldehyde perfusion. Protocol is as follows - Perfusion : perfuse intracardially through the aorta using a pump with the following solutions : solution A (30ml) : 200-300ml/min solution B (500ml) : 200-300ml/min Solution A : cacodylate 0.1M, sodium metabisulfite 10g/l, pH = 6.2 Solution B : cacodylate 0.1M, sodium metabisulfite 10g/l and glutaraldehyde 3-5% pH = 7.5 Post fixation : 15 to 30 min in solution B, then 4 soft washes in Tris 0.05M with sodium metabisulfite 8.5g/l, pH 7.5 (solution C). Tissue sectioning : Cryostat or vibratome sections can be used. Reduction step : Sections are reduced with the solution C containing sodium borohydride (0.1M) for 10 min. Then, the sections are washed 4 times with solution C without sodium borohydride. Application of anti-Dopamine antibodies : The final dilution is 1/2,000 to 1/5,000 in solution C containing triton X100 0.1%, plus 2% of non-specific serum. A dozen sections can be incubated with 2ml of antibody solution overnight at 4°C. Then the sections are washed 3 times (10 min) with solution C. N.B. : Antibodies may be used at a higher dilution. The customer should explore the antibody dilution to reduce the possibility of high background. Note that a substitution in the buffer system as used in our protocol may change the background and the antibody recognition. PAP procedure : Secondary antibody : Sections are incubated with 1/100 dilution of goat anti-rabbit in solution C for 3 hours at 20°C or 1 hour at 37°C. Then washed 3 times (10 min) with solution C. PAP : Sections are incubated with 1/1,000 dilution of rabbit peroxidase anti-peroxidase complex in solution C for 1 hour at 37°C. Then wash 3 times (10 min) with solution C. Detection : Antibody-antigen complexes are revealed using diaminobenzidine (25mg/100ml) (or other chromogen) dissolved in Tris 0.05M and filtrated; 0.05% of H2O2 is added. The sections are incubated for 10 min at 20°C. Reaction is stopped by transfering sections in 5ml of Tris 0.05M.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up