MAB369 Anti-Dopamine Transporter Antibody, NT, clone DAT-Nt

MAB369
100 µL  
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      H, M, Mk, R ICC, IHC, WB R Culture Supernatant Monoclonal Antibody
      Description
      Catalogue NumberMAB369
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      DescriptionAnti-Dopamine Transporter Antibody, NT, clone DAT-Nt
      Alternate Names
      • DAT
      Background InformationThe 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.
      References
      Product Information
      FormatCulture Supernatant
      Control
      • Positive Control: Brain (caudate, putamen, and nucleus accumbens) tissue
      PresentationUnPurified 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.
      Applications
      ApplicationThis Anti-Dopamine Transporter Antibody, N-terminus, clone DAT-Nt is validated for use in IC, IH, WB for the detection of Dopamine Transporter.
      Key Applications
      • Immunocytochemistry
      • Immunohistochemistry
      • Western Blotting
      Application NotesWestern 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).
      Biological Information
      ImmunogenN-terminus of human dopamine transporter fused to Glutathione S-transferase.
      EpitopeN-terminus
      CloneDAT-Nt
      ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
      HostRat
      SpecificityRecognizes 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).
      IsotypeIgG2aκ
      Species Reactivity
      • Human
      • Mouse
      • Monkey
      • Rat
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryThe dopamine transporter (DAT1) mediates the active reuptake of dopamine from the synapse and is a principal regulator of dopaminergic neurotransmission. The DAT1 gene has been implicated in human disorders such as parkinsonism, Tourette syndrome, and substance abuse (Vandenbergh et al., 1992 [PubMed 1359373]).[supplied by OMIM]
      Gene Symbol
      • SLC6A3
      • DAT1
      • ADHD
      • ADD
      • DAT
      Purification MethodUnpurified
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: Q01959 # Amine transporter. Terminates the action of dopamine by its high affinity sodium-dependent reuptake into presynaptic terminals.
      SIZE: 620 amino acids; 68495 Da
      SUBUNIT: Homooligomer; disulfide-linked. Interacts with PRKCABP and TGFB1I1.
      SUBCELLULAR LOCATION: Membrane; Multi-pass membrane protein.
      SIMILARITY: SwissProt: Q01959 ## Belongs to the sodium:neurotransmitter symporter (SNF) family.
      MISCELLANEOUS: This protein is the target of psychomotor stimulants such as amphetamines or cocaine.
      Molecular Weight~ 70-85 kDa
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • 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.
      Storage and Shipping Information
      Storage ConditionsMaintain 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.
      Packaging Information
      Material Size100 µL
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number
      RAT ANTI-DOPAMINE TRANSPORTER2475760
      RAT ANTI-DOPAMINE TRANSPORTER MONOCLONAL ANTIBODY - 21400372140037
      RAT ANTI-DOPAMINE TRANSPORTER MONOCLONAL ANTIBODY - 23910762391076
      RAT ANTI-DOPAMINE TRANSPORTER MONOCLONAL ANTIBODY - 24524952452495
      RAT ANTI-DOPAMINE TRANSPORTER MONOCLONAL ANTIBODY - 20241772024177
      RAT ANTI-DOPAMINE TRANSPORTER - 31886103188610
      RAT ANTI-DOPAMINE TRANSPORTER -25670612567061
      RAT ANTI-DOPAMINE TRANSPORTER -26104752610475
      RAT ANTI-DOPAMINE TRANSPORTER -26644662664466
      RAT ANTI-DOPAMINE TRANSPORTER -27010072701007

      References

      Reference overviewApplicationSpeciesPub Med ID
      Morphological evidence for dopamine interactions with pallidal neurons in primates.
      Eid, L; Parent, M
      Frontiers in neuroanatomy  9  111  2015

      Show Abstract
      26321923 26321923
      Rare autism-associated variants implicate syntaxin 1 (STX1 R26Q) phosphorylation and the dopamine transporter (hDAT R51W) in dopamine neurotransmission and behaviors.
      Cartier, E; Hamilton, PJ; Belovich, AN; Shekar, A; Campbell, NG; Saunders, C; Andreassen, TF; Gether, U; Veenstra-Vanderweele, J; Sutcliffe, JS; Ulery-Reynolds, PG; Erreger, K; Matthies, HJ; Galli, A
      EBioMedicine  2  135-146  2015

      Show Abstract
      Western Blotting25774383 25774383
      Parkin cooperates with GDNF/RET signaling to prevent dopaminergic neuron degeneration.
      Meka, DP; Müller-Rischart, AK; Nidadavolu, P; Mohammadi, B; Motori, E; Ponna, SK; Aboutalebi, H; Bassal, M; Annamneedi, A; Finckh, B; Miesbauer, M; Rotermund, N; Lohr, C; Tatzelt, J; Winklhofer, KF; Kramer, ER
      The Journal of clinical investigation  125  1873-85  2015

      Show Abstract
      25822020 25822020
      Activin A protects midbrain neurons in the 6-hydroxydopamine mouse model of Parkinson's disease.
      Stayte, S; Rentsch, P; Li, KM; Vissel, B
      PloS one  10  e0124325  2015

      Show Abstract
      25902062 25902062
      Are striatal tyrosine hydroxylase interneurons dopaminergic?
      Xenias, HS; Ibáñez-Sandoval, O; Koós, T; Tepper, JM
      The Journal of neuroscience : the official journal of the Society for Neuroscience  35  6584-99  2015

      Show Abstract
      25904808 25904808
      Comprehensive functional characterization of murine infantile Batten disease including Parkinson-like behavior and dopaminergic markers.
      Dearborn, JT; Harmon, SK; Fowler, SC; O'Malley, KL; Taylor, GT; Sands, MS; Wozniak, DF
      Scientific reports  5  12752  2015

      Show Abstract
      26238334 26238334
      Genetic variation in COMT activity impacts learning and dopamine release capacity in the striatum.
      Simpson, EH; Morud, J; Winiger, V; Biezonski, D; Zhu, JP; Bach, ME; Malleret, G; Polan, HJ; Ng-Evans, S; Phillips, PE; Kellendonk, C; Kandel, ER
      Learning & memory (Cold Spring Harbor, N.Y.)  21  205-14  2014

      Show Abstract
      24639487 24639487
      Recombineering strategies for developing next generation BAC transgenic tools for optogenetics and beyond.
      Ting, JT; Feng, G
      Frontiers in behavioral neuroscience  8  111  2014

      Show Abstract
      24772073 24772073
      Axonal targeting of the serotonin transporter in cultured rat dorsal raphe neurons is specified by SEC24C-dependent export from the endoplasmic reticulum.
      Montgomery, TR; Steinkellner, T; Sucic, S; Koban, F; Schüchner, S; Ogris, E; Sitte, HH; Freissmuth, M
      The Journal of neuroscience : the official journal of the Society for Neuroscience  34  6344-51  2014

      Show Abstract
      24790205 24790205
      Progranulin gene delivery protects dopaminergic neurons in a mouse model of Parkinson's disease.
      Van Kampen, JM; Baranowski, D; Kay, DG
      PloS one  9  e97032  2014

      Show Abstract
      ImmunohistochemistryMouse24804730 24804730

      Data Sheet

      Title
      RAT ANTI-DOPAMINE TRANSPORTER MONOCLONAL ANTIBODY

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