Overview

  • Product name
  • Description
    Rabbit polyclonal to Dopamine
  • Host species
    Rabbit
  • Specificity
    ab8888 targets conjugated dopamine, it does not recognize free dopamine.
  • Tested applications
    Suitable for: IHC-P, IHC-Fr, ICCmore details
  • Immunogen

    Chemical/ Small Molecule corresponding to Dopamine conjugated to Bovine Serum Albumin (BSA).

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Constituents: ddH20, 50% Glycerol
  • Purity
    Whole antiserum
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab8888 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
IHC-P 1/1000 - 1/5000. Fixation of tissue for use with this antibody should be done with gluteraldehyde. The use of paraformaldehyde in conjunction with gluteraldehyde may improve staining results.
IHC-Fr Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.

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

  • ab8888 at 1/500 staining human adrenal tissue sections by IHC-P. The tissue was formaldehyde fixed and blocked with serum before incubation with the antibody for 30 minutes at 22°C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.

    See Abreview

References

This product has been referenced in:
See all 5 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Mouse midbrain)
Specification
Mouse midbrain
Fixative
5% Glutaraldehyde
Blocking step
Serum as blocking agent for 6 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Aug 10 2012

Answer

Thank you for confirming this information and for your help and cooperation with this case.

As requested, I have asked our accounting department to issue you with a credit note. This can then be redeemed against the invoice of a future order.

Credit ID: #####

As usual if you have any further questions regarding this credit note, please contact the accounts department by email at creditcontrol@abcam.com. Please refer to the credit ID number in any correspondence with the accounting department.

I would like to wish the customer good luck with their research. The technical team is always at your service, should you require further expert advice.

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Question

Dear technical support team:

This customer has purchased ab8888 (Anti-Dopamine antibody)and has conducted the IHC several times with mouse sample. The results show on signal, therefore, he would like to ask for your help to modify his experiment step, could you please offer any suggestion to improve his result?

I attached the image in this letter and his experiment step as follow:


1) Abcam product code: ab8888

2) lot number gr57936-1

3) Description of the problem: No signal

4) Sample preparation: Species mouse

Type of sample: perfusion fixed frozen sections:

Sample preparation

Positive control: WT mice

Negative control: DDC KI mice



5) Fixation step

Yes

If yes: Fixative agent and concentration:solution B (Abecam ab8888 protocol)

Fixation time:overnight

Fixation temperature: 4 c



6) Antigen retrieval method

Your protocol does not have retrival

7) Permeabilization method: Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?

Permeabilizing agent and concentration:

OCT emded in dry ice)



8) Blocking agent (eg BSA, serum…): BSA

Concentration:2.5%

Blocking time: Abecam ab8888 protocol

Blocking temperature :RT(Abecam ab8888 protocol)



9) Endogenous peroxidases blocked?

Endogenous biotins blocked?

0.3% H2O2

10) Primary antibody (If more than one was used, describe in “additional notes”) :

Concentration or dilution:1:500

Diluent buffer : blocking buffer

Incubation time: :overnight



11) Secondary antibody:biogenex IHC kit

Species:

Reacts against: :biogenex IHC kit

Concentration or dilution :biogenex IHC kit

Diluent buffer :biogenex IHC kit

Incubation time :biogenex IHC kit

enzyme conjugate: :biogenex IHC kit



12) Washing after primary and secondary antibodies:

Buffer

Number of washes 3



13) Detection method :DAB



14) How many times have you run this staining? 3

Do you obtain the same results every time? YES

What steps have you altered to try and optimize the use of this antibody?

Each time uses your Abecam ab8888 protocol

2 nd ab use biogenex IHC kit(1 month ago,I asked to your department,)

Could you please help this customer to solve the problem?

Thanks for your kindly help

Best regards

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Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

I would like to reassure you that ab8888 is tested and covered by our 6 month guarantee for use inIHC-Fr and IHC-P. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

Reviewing this case, I would like to offer some suggestions to help optimize the results from ab8888. I would also appreciate if you can confirm some further details:

1. I am sorry this antibody is not tested in perfusion fixed tissue (IHC-FoFr), but we can make an exception in this case as its similar to IHC-Fr which has been tested.

2. Please confirm what mouse tissue has been tested. Was this brain tissue? which part of the brain?

3. Could you confirm if the overnight incubation with primary antibody at 4oC? This lower temperature can often provide more efficient and specific staining.

4. To increase the signal, I can recommend to try a higher concentration of antibody, 1:200. This can sometimes require optimization and dilutions on protocols are a guideline only.

5. I can suggest to try including antigen retrievalas this may need optimization by the end user.
As it has been incubated in PFA overnight to fix, I can suggest this may help expose the antigen.

6. Is the secondary antibody anddetection Biogenex kit working well with other primary antibodies? Is it suitable for detection of rabbit polyclonalor IgG primary? I would appreciate if you are able to provide the product code. What secondary antibody does this contain?

7. Could you confirm what is in the antibody dilution buffer and wash buffer? I can recommend to use PBS containing 0.2% Tween 20. This will help to solubilize the antibody.

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

Read More

Answer

Thank you for contacting us.

You are quite correct. because of its size, the dopamine need to be attached to a macromolecule to be effectively visualized.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for contacting us.

If you are using a Rabbit primary antibody in Mouse sections, the recommended secondary must be an anti-Rabbit raised preferably in a species which is as phylogenetically different as possible (distant) from mouse species to avoid cross reactivity.

We have some secondaries in our catalogue raised in Goat that could be appropriate for your assay:

https://www.abcam.com/Rabbit-IgG-secondary-antibody-F-ab-2-F-ab-2-Fragment-ab6112.html

https://www.abcam.com/Goat-F-ab-2-polyclonal-Secondary-Antibody-to-Rabbit-IgG-H-L-Biotin-pre-adsorbed-ab6012.html

https://www.abcam.com/Rabbit-IgG-secondary-antibody-F-c-F-ab-2-Fragment-ab6021.html

https://www.abcam.com/Rabbit-IgG-secondary-antibody-H-L-F-ab-2-Fragment-ab6013.html

https://www.abcam.com/Rabbit-IgG-secondary-antibody-H-L-Fab-Fragment-ab7171.html

It is also very important to use a blocking agent such as serum from the same species as the secondary was raised in. If you used any of the secondaries suggested, therefore, 10% goat serum would be the most suitable blocking agent.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information

Read More

Answer

Thank you for contacting us. I can confirm this antibody is suitable for conducting WB in mouse samples as the target (Dopamine) is a small molecule that does not vary between species. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

I have been contacted by the origonator of this product who has some additional information that you may be interested in. I have been contacted by the originator of this product who has some additional information that you may be interested in. Perfusion protocol for Adult male Sprague Dawley (weight around 0.5 kg) : 1-The animals can be deeply anaesthetized for example with urethane (0.5-1.5g/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 0.1 M phosphate-buffer (PB), pH 7.2, (two minutes). b) 600 ml of cold 4% paraformaldehyde and 2% glutaraldehyde in 0.1 M phosphate-buffer (PB), pH 7.2, (ten minutes). c) Dissect out the brains and place in a solution of 4% paraformaldehyde in 0.1M PB, pH 7.2, at 4ºC for twelve to sixteen hours. d) Before the brains will be cut on a freezing microtome, we must include the brain in growing concentrations of sucrose (a first bain of 5% of sucrose in PBS until the brains sank), after that we will repeat the same process in a solution with a higher level of sucrose (10%), 20%, 25% and finally 30%.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (adrenal)
Specification
adrenal
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer pH 6.0
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Oct 05 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Adrenal)
Specification
Adrenal
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Oct 05 2007

1-10 of 11 Abreviews or Q&A

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

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