MAB369
Anti-Dopamine Transporter Antibody, NT, clone DAT-Nt
culture supernatant, clone DAT-Nt, Chemicon®
Synonym(s):
DAT
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All Photos(4)
About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41
Recommended Products
biological source
rat
Quality Level
antibody form
culture supernatant
antibody product type
primary antibodies
clone
DAT-Nt, monoclonal
species reactivity
rat, human, mouse, monkey
manufacturer/tradename
Chemicon®
technique(s)
immunocytochemistry: suitable
immunohistochemistry: suitable
western blot: suitable
isotype
IgG2aκ
NCBI accession no.
UniProt accession no.
shipped in
dry ice
target post-translational modification
unmodified
Gene Information
human ... SLC6A3(6531)
General description
The transmembrane dopamine transporter (DAT) is located on the presynaptic nerve terminal and is responsible for terminating dopaminergic transmission by transporting dopamine from the synaptic cleft into the dopaminergic neuron (reuptake). Dopaminergic pathways have been strongly implicated in reward and addiction, motivation, alcoholism, ADHD and degenerative motor diseases such as Parkinson′s, Huntington′s and Chorea.
Specificity
Recognizes Dopamine transporter, N-terminus. Shows no cross reactivity to the closely related serotonin and norepinephrine transporters (Miller, 1997). Immunolocalization of DAT on paraformaldehyde fixed frozen sections of human brain using MAB369 shows dense punctate staining throughout the caudate, putamen and accumbens (Miller, 1997).
Immunogen
Epitope: N-terminus
N-terminus of human dopamine transporter fused to Glutathione S-transferase.
Application
Research Category
Neuroscience
Neuroscience
Research Sub Category
Ion Channels & Transporters
Ion Channels & Transporters
This Anti-Dopamine Transporter Antibody, N-terminus, clone DAT-Nt is validated for use in IC, IH, WB for the detection of Dopamine Transporter.
Western blot: Recognizes a diffuse band at approximately 70-85 kDa on western blots of extracts (20 ug total protein) from human caudate, putamen, and nucleus accumbens.
Immunohistochemistry: 4% paraformaldehyde fixed tissue (care should be taken not to over-fix tissue); perfusion followed by less than 90 minutes post-fixation, then cryoprotect. Suggested working dilution 1:1,000-1:10,000. If using on rat tissue, absorbed anti-rat secondary antibodies are recommended, and the use of rat PAP systems, or ABC detection will enhance sensitivity.
Immunocytochemistry: 1:5,000 to 1:10,000.
Immunoblotting (not recommended for use on rat)
Optimal working dilutions and protocols must be determined by end user.
IMMUNOHISTOCHEMISTRY PROTOCOL FOR MAB369
This antibody has been used successfully on 30 μm, free floating, 4% paraformaldehyde fixed mouse brain tissue. All steps are performed under constant agitation. Suggested protocol follows.
1) 3 x 10 minute washes in TBS (with or without 0.25% Triton).
2) Incubate for 30 minutes in TBS with 3% serum (same as host from secondary antibody).
3) Incubate primary antibody diluted appropriately in TBS with 1% serum (same as host from secondary antibody) (with or without 0.25% Triton) for 2 hours at room temperature followed by 16 hours at 4°C.
4) 3 x 10 minute washes in TBS.
5) Incubate with secondary antibody diluted appropriately in TBS with 1% serum (same as host from secondary antibody).
6) 3 x 10 minute washes in TBS.
7) ABC Elite (1:200 Vector Labs) in TBS.
8) 2 x 10 minute washes in TBS.
9) 1 x 10 minute wash in phosphate buffer (no saline).
10) DAB reaction with 0.06% NiCl added for intensification.
11) 2 x 10 minute washes in PBS.
12) 1 x 10 minute wash in phosphate buffer (no saline).
Immunohistochemistry: 4% paraformaldehyde fixed tissue (care should be taken not to over-fix tissue); perfusion followed by less than 90 minutes post-fixation, then cryoprotect. Suggested working dilution 1:1,000-1:10,000. If using on rat tissue, absorbed anti-rat secondary antibodies are recommended, and the use of rat PAP systems, or ABC detection will enhance sensitivity.
Immunocytochemistry: 1:5,000 to 1:10,000.
Immunoblotting (not recommended for use on rat)
Optimal working dilutions and protocols must be determined by end user.
IMMUNOHISTOCHEMISTRY PROTOCOL FOR MAB369
This antibody has been used successfully on 30 μm, free floating, 4% paraformaldehyde fixed mouse brain tissue. All steps are performed under constant agitation. Suggested protocol follows.
1) 3 x 10 minute washes in TBS (with or without 0.25% Triton).
2) Incubate for 30 minutes in TBS with 3% serum (same as host from secondary antibody).
3) Incubate primary antibody diluted appropriately in TBS with 1% serum (same as host from secondary antibody) (with or without 0.25% Triton) for 2 hours at room temperature followed by 16 hours at 4°C.
4) 3 x 10 minute washes in TBS.
5) Incubate with secondary antibody diluted appropriately in TBS with 1% serum (same as host from secondary antibody).
6) 3 x 10 minute washes in TBS.
7) ABC Elite (1:200 Vector Labs) in TBS.
8) 2 x 10 minute washes in TBS.
9) 1 x 10 minute wash in phosphate buffer (no saline).
10) DAB reaction with 0.06% NiCl added for intensification.
11) 2 x 10 minute washes in PBS.
12) 1 x 10 minute wash in phosphate buffer (no saline).
Target description
~ 70-85 kDa
Physical form
UnPurified tissue culture supernatant, filtered through a 0.2 μm membrane prior to vialing. Product contains 20%FBS and Ciprofloxacin at final concentration of 10μg/mL.
Unpurified
Storage and Stability
Maintain for 1 year at -20°C from date of shipment. Aliquot to avoid repeated freezing and thawing. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Analysis Note
Control
Positive Control: Brain (caudate, putamen, and nucleus accumbens) tissue
Positive Control: Brain (caudate, putamen, and nucleus accumbens) tissue
Legal Information
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
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recommended
Storage Class Code
10 - Combustible liquids
WGK
WGK 1
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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Weimin C Hong et al.
FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 27(8), 2995-3007 (2013-04-25)
The dopamine transporter (DAT) clears the extracellular dopamine released during neurotransmission and is a major target for both therapeutic and addictive psychostimulant amphetamines. Amphetamine exposure or activation of protein kinase C (PKC) by the phorbol ester PMA has been shown
Flotillins regulate membrane mobility of the dopamine transporter but are not required for its protein kinase C dependent endocytosis.
Sorkina, T; Caltagarone, J; Sorkin, A
Traffic null
Methamphetamine increases locomotion and dopamine transporter activity in dopamine d5 receptor-deficient mice.
Hayashizaki, S; Hirai, S; Ito, Y; Honda, Y; Arime, Y; Sora, I; Okado, H; Kodama, T; Takada, M
Testing null
D Tomas et al.
PloS one, 11(4), e0153424-e0153424 (2016-04-15)
The dyskinesia of Parkinson's Disease is most likely due to excess levels of dopamine in the striatum. The mechanism may be due to aberrant synthesis but also, a deficiency or absence of the Dopamine Transporter. In this study we have
Armando G Salinas et al.
Neuropharmacology, 108, 275-283 (2016-04-03)
The striatum is typically classified according to its major output pathways, which consist of dopamine D1 and D2 receptor-expressing neurons. The striatum is also divided into striosome and matrix compartments, based on the differential expression of a number of proteins
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