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
  • 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:
  • Banks DA  et al. MK-STYX Alters the Morphology of Primary Neurons, and Outgrowths in MK-STYX Overexpressing PC-12 Cells Develop a Neuronal Phenotype. Front Mol Biosci 4:76 (2017). ICC/IF ; Rat . Read more (PubMed: 29250526) »
  • Wu SF  et al. Dopamine modulates hemocyte phagocytosis via a D1-like receptor in the rice stem borer, Chilo suppressalis. Sci Rep 5:12247 (2015). Read more (PubMed: 26179416) »
See all 3 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

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Abcam guarantees this product to work in the species/application used in this Abreview.
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.

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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.

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Question
Answer

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

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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.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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