Overview

  • Product name
    Anti-Dopamine Transporter antibody [hDAT-NT]
    See all Dopamine Transporter primary antibodies
  • Description
    Rat monoclonal [hDAT-NT] to Dopamine Transporter
  • Host species
    Rat
  • Specificity
    This antibody recognizes the N-terminus of the dopamine transporter (DAT-NT) in mice and humans.
  • Tested applications
    Suitable for: IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Fusion protein corresponding to Dopamine Transporter aa 1-66 (N terminal).

  • Positive control
    • IHC-P: Mouse brain tissue.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.4
    Constituents: PBS, 0.81% Sodium chloride, 0.16% Sodium phosphate, 0.02% Potassium chloride, 0.04% Potassium phosphate
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    hDAT-NT
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab5990 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. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

Target

  • Function
    Amine transporter. Terminates the action of dopamine by its high affinity sodium-dependent reuptake into presynaptic terminals.
  • Involvement in disease
    Defects in SLC6A3 are the cause of dystonia-parkinsonism infantile (DYTPRI) [MIM:613135]. It is a neurodegenerative disorder characterized by infantile onset of parkinsonism and dystonia. Other neurologic features include global developmental delay, bradikinesia and pyramidal tract signs.
  • Sequence similarities
    Belongs to the sodium:neurotransmitter symporter (SNF) (TC 2.A.22) family. SLC6A3 subfamily.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • DA transporter antibody
    • DAT 1 antibody
    • DAT antibody
    • DAT1 antibody
    • Dopamine transporter 1 antibody
    • Dopamine transporter antibody
    • PKDYS antibody
    • SC6A3_HUMAN antibody
    • SLC6A3 antibody
    • Sodium dependent dopamine transporter antibody
    • Sodium-dependent dopamine transporter antibody
    • Solute carrier family 6 (neurotransmitter transporter dopamine), member 3 antibody
    • Solute carrier family 6 (neurotransmitter transporter), member 3 antibody
    • Solute carrier family 6 member 3 antibody
    • Variable number tandem repeat (VNTR) antibody
    see all

Images

  • ab5990 staining of dopamine transporter in mouse brain (substantia nigra) tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Heat-mediated antigen retrieval was carried out using citric acid. Samples were incubated with primary antibody (1/1000) for two hours at room temperature. A Biotin-conjugated goat anti-rat IgG polyclonal was used as the secondary antibody.

References

This product has been referenced in:
  • Playne R  et al. Generation of dopamine neuronal-like cells from induced neural precursors derived from adult human cells by non-viral expression of lineage factors. J Stem Cells Regen Med 14:34-44 (2018). Read more (PubMed: 30018471) »
  • Arima Y  et al. Brain micro-inflammation at specific vessels dysregulates organ-homeostasis via the activation of a new neural circuit. Elife 6:N/A (2017). Read more (PubMed: 28809157) »
See all 11 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
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 May 16 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Antigen retrieval step
None
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Permeabilization
No
Fixative
Paraformaldehyde

Dr. Karine Thibault

Verified customer

Submitted Aug 19 2013

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (rat dopaminergic neuron)
Specification
rat dopaminergic neuron
Fixative
Formaldehyde
Permeabilization
No
Blocking step
saponin as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Apr 05 2013

Answer

Thank you for contacting us.

ab104224
DISCOUNT CODE: xxxxxxxx
Expiration date: xxxxxxxxxx

I am very pleased to hear that your customer would like to accept our offer and test ab104224 in IHC-Fr. 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.


ab5990
DISCOUNT CODE: xxxxxxxxx
Expiration date: xxxxxxxxxxx

I am very pleased to hear that your customer would like to accept our offer and test ab5990 in IF. To redeem this offer, please submit an Abreview for IF 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.

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

Read More

Question

Pasted you can find the questionnaire you sent us. We look forward to hearing from you soon.
1) Abcam product code ab
ab5990

2) Abcam order reference number or product batch number

3) Description of the problem
The antibody does not detect the predicted band for DAT (74 KDa)

4) Sample preparation:
Type of sample (whole cell lysates, fraction, recombinant protein…): Brain tissue (N. Accumbens) homogenate
Lysis buffer
Protease inhibitors: Roche
Phosphatase inhibitors: Calbiochem
Reducing agent: 2-mercaptoethanol
Boiling for ≥5 min: yes
Protein loaded ug/lane or cells/lane: 40-60 ug protein/lane
Positive control
Negative control

5) Percentage of gel: 10%
Type of membrane: PVDF
Protein transfer verified: Yes
Blocking agent and concentration: 1% BSA in TBS-Tween 1,5%
Blocking time: > 1h
Blocking temperature: Room Temperature

6) Primary antibody (If more than one was used, describe in “additional notes”) :
Concentration or dilution: 1:500 – 1:300
Diluent buffer: TBS-Tween 1,5%
Incubation time: 20 h aprox.
Incubation temperature: 1-2 h Room Temperature - 4º C overnight

7) Secondary antibody:
Species: goat anti-rat
Reacts against: rat IgG
Concentration or dilution: 1:5000 – 1:2000
Diluent buffer: TBS-Tween 1,5%
Incubation time: 1h
Incubation temperature: Room Temperature
Fluorochrome or enzyme conjugate: HRP

8) Washing after primary and secondary antibodies:
Buffer: TBS-Tween 1,5%
Number of washes: 3 (10 min each wash)

9) Detection method: Chemifluorescence
10) How many times have you run this staining?
Do you obtain the same results every time? Yes
What steps have you altered to try and optimize the use of this antibody?
We have changed the ug Protein/lane, we have increased primary and secondary antibodies concentrations.

Read More
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 ab5990 is tested and covered by our 6 month guarantee for use in WB and mouse, rat and human samples. 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 optimise the results from ab5990.

1.) I suggest to decrease the boiling temperature to 65C and to prolong the heating time up to 10 min. Your target is a multi-pass membrane protein, and these proteins have the unfortunate tendency to aggregate during boiling periods with high temperatures and beta- mercaptoethanol due to their hydrophobic nature.

2.) I also recommend to reduce the amount of Tween to 0.1%. Too much tween will lead to reduced binding of the antibody since the tween is acting as detergent.

I would also appreciate if you can confirm some further details:

Is the secondary antibody working with other primary antibodies?

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 your enquiry,.

We recommend that antibodies should be stable for at least 1 year at the recommended storage temperature. This can be much longer for frozen antibodies. However, please noteour Abpromise guarantee is provided for 6 months after purchase.

With regards to concentration and binding to the tube when freezing, the best way to avoid this is to ensure aliquots are no less than 10 ul and ensure the lid on the tube is pressed well closed. Smaller volumes can evaporate and dry out easily, altering concentration as well as product quality.

I hope this information will be helpful to you. Should you have any further questions, please do not hesitate to contact us.

Read More

Answer

Thank you for contacting us.  We have three Dopamine Transporter antibodies that will be guaranteed for use in WB on rat samples: ab5981, ab5990, and ab18548.  Please find their datasheets attached.  I would be happy to help you further if you have any additional questions about our products. 

Read More

Answer

Thank you for your enquiry. I regret to inform you that blocking peptides are not available for these 3 antibodies. If you have any additional questions, please don't hesitate to contact us again.

Read More

Answer

Thank you for your enquiry. Are looking for a dopamine transporter antibody or a dopamine antibody? You indicated ab5990 - Rat monoclonal to Dopamine Transporter - which is suitable for use in ICC but has not yet been tested for cross-reactivity with species other than human and rat. Ab5981 is another suggestion; it is a rat monoclonal [hDAT-LOOP] to Dopamine Transporter and works in ICC, although has not yet been tested for cross-reactivity with species other than human and rat. We do not routinely offer free or trial sized samples for testing purposes. Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement. Should you decide to go ahead and purchase this product, please let us know how you get on by submitting an Abreview and in return we will offer you 50 Abcam Points which can be redeemed on a number of rewards (a further 100 points will be offered for an image). If you have any additional questions, please contact us again.

Read More

Answer

I'm sorry to hear you are having a problem with ab5990. Rat DAT contains various carbohydrate moieties and neuramnidase and N-Glycanase treatment reduces it size to ~55kD, so I am wondering if this could be the reason why you see a lower MW band. This antibody has been used successfully for Western blot and has been cited in several publications on the datasheet such as Hersch SM et al. Subcellular localization and molecular topology of the dopamine transporter in the striatum and substantia nigra. J Comp Neurol 388:211-27 (1997). PubMed: 9368838. In this paper, for example, the authors loaded 100 ug of membranes from rat striatum, blocked in 5% milk for 1 hr at room temp, incubated at 4 degrees overnight at 1:500 in blocking buffer, and incubated in secondary at 1:10,000 for 1 hour and detected a band of approximately 90 kDa. I am wondering if any of these steps can be included in your current protocol to improve the detection of the correct size band. We sell two other DAT antibodies, Ab5981 to the second extracellular loop and cited in the paper above, and ab18548 which has not been cited but reacts with a broad 80 kDa band that can be competed with the immunizing peptide. Please let me know if this helps and do not hesitate to contact us for further advice,

Read More

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

Sign up