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    dopamine-transporter-antibody-epr19695-ab184451.pdf

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Neuroscience Neurotransmitter Biogenic Amines Dopamine
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RecombinantRabMAb

Recombinant Anti-Dopamine Transporter antibody [EPR19695] (ab184451)

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  • Certificate of Compliance
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Western blot - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
  • Western blot - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
  • Immunohistochemistry (Frozen sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
  • Immunohistochemistry (Frozen sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
  • Immunohistochemistry (Frozen sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
  • Immunohistochemistry (Frozen sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
  • Immunohistochemistry (Frozen sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
  • Immunoprecipitation - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
  • Anti-Dopamine Transporter antibody [EPR19695] (ab184451)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19695] to Dopamine Transporter
  • Suitable for: WB, IHC-P, IHC-Fr, IP
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free

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Overview

  • Product name

    Anti-Dopamine Transporter antibody [EPR19695]
    See all Dopamine Transporter primary antibodies
  • Description

    Rabbit monoclonal [EPR19695] to Dopamine Transporter
  • Host species

    Rabbit
  • Specificity

    Human species is recommended based on IHC-Fr results, we do not guarantee
    WB and IHC-P for human.
  • Tested applications

    Suitable for: WB, IHC-P, IHC-Fr, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Unboiled Mouse and rat striatum lysates. IHC-P: Mouse and rat striatum tissues IHC-Fr: Mouse brain (Coronal section), rat brain (sagittal section), normal human striatum, normal human substantia nigra. IP: Mouse striatum whole cell lysate.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR19695
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurotransmitter
    • Biogenic Amines
    • Dopamine
    • Neuroscience
    • Neurotransmitter
    • Transporters
    • Dopamine
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-Dopamine Transporter antibody [EPR19695] - BSA and Azide free (ab221845)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human Dopamine Transporter protein (ab112352)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab184451 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
WB
1/1000. Detects a band of approximately 70-85 kDa (predicted molecular weight: 69 kDa).
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr
1/500.
IP
1/30.
Notes
WB
1/1000. Detects a band of approximately 70-85 kDa (predicted molecular weight: 69 kDa).
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr
1/500.
IP
1/30.

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.
  • Target information above from: UniProt accession Q01959 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 6531 Human
    • Entrez Gene: 13162 Mouse
    • Entrez Gene: 24898 Rat
    • Omim: 126455 Human
    • SwissProt: Q01959 Human
    • SwissProt: Q61327 Mouse
    • SwissProt: P23977 Rat
    • Unigene: 406 Human
    • Unigene: 41993 Mouse
    • Unigene: 10093 Rat
    see all
  • 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

  • Western blot - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Western blot - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    All lanes : Anti-Dopamine Transporter antibody [EPR19695] (ab184451) at 1/1000 dilution

    Lane 1 : Mouse striatum lysate boiled
    Lane 2 : Mouse striatum lysate unboiled
    Lane 3 : Rat striatum lysate boiled
    Lane 4 : Rat striatum lysate unboiled

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 69 kDa



    We recommend not to boil the samples after lysis to get desired WB results

  • Western blot - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Western blot - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    All lanes : Anti-Dopamine Transporter antibody [EPR19695] (ab184451) at 1/1000 dilution

    Lane 1 : Mouse striatum lysate
    Lane 2 : Rat striatum lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Rat liver lysate
    Lane 5 : Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate
    Lane 6 : C6 (Rat glial tumor cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 69 kDa
    Observed band size: 70-85 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Dopamine transporters are distributed widely across the brain in regions with high dopaminergic activity, such as the striatum and substantia nigra (PMID:1765147), so non-related tissues and cells were chosen as negative controls; and DAT is a glycoprotein with molecular weight of 70-85KD (PMID:25356398, PMID: 20643191).

  • Immunohistochemistry (Frozen sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Immunohistochemistry (Frozen sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)

    IHC image of Dopamine Transporter staining in a section of formalin fixed paraffin embedded normal human striatum* performed on a Leica BOND™.  The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab184451, 5ugml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen.

    The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Immunohistochemistry (Frozen sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Immunohistochemistry (Frozen sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)

    IHC image of Dopamine Transporter staining in a section of formalin fixed paraffin embedded normal human substantia nigra* performed on a Leica BOND™.  The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was incubated with ab184451, 5ugml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Immunohistochemistry (Frozen sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Immunohistochemistry (Frozen sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)

    Negative control image: IHC image of Dopamine Transporter staining in a section of formalin fixed paraffin embedded normal human liver* performed on a Leica BOND™.  The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins..The section was incubated with ab184451, 5ugml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Immunohistochemistry (Frozen sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Immunohistochemistry (Frozen sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen Rat brain (sagittal section) tissue labeling Dopamine Transporter with ab184451 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
    The result showed cytoplasmic staining on rat striatum.
    The nuclear counterstain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 at 1/1000 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Immunohistochemical analysis of paraffin-embedded Mouse striatum tissue labeling Dopamine Transporter with ab184451 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
    Membrane and cytoplasm staining on mouse striatum is observed.
    Counter stained with Hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Dopamine Transporter with ab184451 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
    Counter stained with Hematoxylin.
     
    Negative control: Negative staining on mouse liver.
    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Immunohistochemical analysis of paraffin-embedded Rat striatum tissue labeling Dopamine Transporter with ab184451 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
    Cytoplasm staining on rat striatum is observed.
    Counter stained with Hematoxylin. 

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling Dopamine Transporter with ab184451 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
    Counter stained with Hematoxylin.
     
    Negative control: Negative staining on rat spleen.
    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Immunohistochemistry (Frozen sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Immunohistochemistry (Frozen sections) - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen Mouse brain (Coronal section) tissue labeling Dopamine Transporter with ab184451 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
    The result showed cytoplasmic staining on mouse striatum but negative on lateral ventricle (LA).
    The nuclear counterstain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 at 1/1000 dilution.

  • Immunoprecipitation - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Immunoprecipitation - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)

    Dopamine Transporter was immunoprecipitated from 0.35mg of Mouse striatum whole cell lysate with ab184451 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using ab184451 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: Mouse striatum whole cell lysate, 10µg (Input).

    Lane 2: ab184451 IP in Mouse striatum whole cell lysate.

    Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab184451 in Mouse striatum whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 30 seconds.

  • Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
    Anti-Dopamine Transporter antibody [EPR19695] (ab184451)

Protocols

  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (9)

Publishing research using ab184451? Please let us know so that we can cite the reference in this datasheet.

ab184451 has been referenced in 9 publications.

  • Jasmin M  et al. Netrin-1 and its receptor DCC modulate survival and death of dopamine neurons and Parkinson's disease features. EMBO J 40:e105537 (2021). PubMed: 33351190
  • Yamane M  et al. Hair-Follicle-Associated Pluripotent (HAP) Stem Cells Can Extensively Differentiate to Tyrosine-Hydroxylase-Expressing Dopamine-Secreting Neurons. Cells 10:N/A (2021). PubMed: 33920157
  • Li S  et al. Multiregional profiling of the brain transmembrane proteome uncovers novel regulators of depression. Sci Adv 7:N/A (2021). PubMed: 34290087
  • Williams MT  et al. Whole brain proton irradiation in adult Sprague Dawley rats produces dose dependent and non-dependent cognitive, behavioral, and dopaminergic effects. Sci Rep 10:21584 (2020). PubMed: 33299021
  • Raffaelli FM  et al. Dopamine receptor D1- and D2-agonists do not spark brown adipose tissue thermogenesis in mice. Sci Rep 10:20203 (2020). PubMed: 33214601
  • Li W  et al. Effects of Combined Bushen Zhichan Recipe and Levodopa in a Rodent Model of Parkinson Disease: Potential Mechanisms. Med Sci Monit 26:e922345 (2020). PubMed: 32555131
  • Wu J  et al. Distinct Connectivity and Functionality of Aldehyde Dehydrogenase 1a1-Positive Nigrostriatal Dopaminergic Neurons in Motor Learning. Cell Rep 28:1167-1181.e7 (2019). PubMed: 31365862
  • Wang J  et al. MicroRNA-124 Enhances Dopamine Receptor Expression and Neuronal Proliferation in Mouse Models of Parkinson's Disease via the Hedgehog Signaling Pathway by Targeting EDN2. Neuroimmunomodulation 26:174-187 (2019). PubMed: 31454817
  • Regan SL  et al. Knockout of latrophilin-3 in Sprague-Dawley rats causes hyperactivity, hyper-reactivity, under-response to amphetamine, and disrupted dopamine markers. Neurobiol Dis 130:104494 (2019). PubMed: 31176715

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