RecombinantRabMAb

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

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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 (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

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

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

  • Description

    Rabbit monoclonal [EPR19695] to Dopamine Transporter

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, IHC-Fr, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat

  • 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) and rat brain (sagittal section) tissues. 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

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.
  • 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

    • 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)
      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)

    References (4)

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

    ab184451 has been referenced in 4 publications.

    • 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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