Anti-Dopamine Transporter antibody [EPR19695] (ab184451)

Anti-Dopamine Transporter antibody [EPR19695] ab184451 is a rabbit monoclonal antibody that is used in Dopamine Transporter western blotting and IHC. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone EPR19695 is cited in over 30 publications

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Images

Western blot - Anti-Dopamine Transporter antibody [EPR19695] (AB184451), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	IHC-Fr	IHC-P	ICC
Human	Expected	Expected	Expected	Tested	Not recommended
Mouse	Tested	Tested	Tested	Tested	Not recommended
Rat	Expected	Tested	Tested	Tested	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/30	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Rat, Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/1000	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes -
Species Rat	Dilution info 1/500	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1/250	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/250	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Rat, Mouse	Dilution info -	Notes -

Associated Products

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Target data

See full target information SLC6A3

Function

Mediates sodium- and chloride-dependent transport of dopamine (PubMed:10375632, PubMed:11093780, PubMed:1406597, PubMed:15505207, PubMed:19478460, PubMed:8302271). Also mediates sodium- and chloride-dependent transport of norepinephrine (also known as noradrenaline) (By similarity). Regulator of light-dependent retinal hyaloid vessel regression, downstream of OPN5 signaling (By similarity).

Alternative names

DAT1, SLC6A3, Sodium-dependent dopamine transporter, DA transporter, DAT, Solute carrier family 6 member 3

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR19695
Purification technique: Affinity purification Protein A
Specificity: Human species is recommended based on IHC-P results, we do not guarantee IHC-Fr, WB and IP for human.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

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

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 batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

12 product images

Western blot - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
We recommend not to boil the samples after lysis to get desired WB results.
Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as a GAPDH loading control.
All lanes: Western blot - Anti-Dopamine Transporter antibody [EPR19695] (ab184451) at 1/1000 dilution
Lane 1: Mouse striatum lysate boiled at 20 µg
Lane 2: Mouse striatum lysate unboiled at 20 µg
Lane 3: Rat striatum lysate boiled at 20 µg
Lane 4: Rat striatum lysate unboiled at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 68 kDa
Western blot - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
Blocking/Dilution buffer: 5% NFDM/TBST.
Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as GAPDH loading control.
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).
All lanes: Western blot - Anti-Dopamine Transporter antibody [EPR19695] (ab184451) at 1/1000 dilution
Lane 1: Mouse striatum lysate at 20 µg
Lane 2: Rat striatum lysate at 20 µg
Lane 3: Mouse kidney lysate at 20 µg
Lane 4: Rat liver lysate at 20 µg
Lane 5: Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate at 20 µg
Lane 6: C6 (Rat glial tumor cell line) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 68 kDa
Observed band size: 70-85 kDa
Exposure time: 3min
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)
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 (Formalin/PFA-fixed paraffin-embedded 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)
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) (Goat Anti-Rabbit IgG H&L (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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/1000 dilution.
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) (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 Goat Anti-Rabbit IgG H&L (HRP) 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)
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) (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.
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) (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 (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.
All lanes: Immunoprecipitation - Anti-Dopamine Transporter antibody [EPR19695] (ab184451)
Predicted band size: 68 kDa
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) (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 Goat Anti-Rabbit IgG H&L (HRP) 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)
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) (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)
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) (Goat Anti-Rabbit IgG H&L (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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/1000 dilution.