Ordering Information
- : MAB369
- : 100 μL
- : Variable
Product Images
Rat anti-Dopamine Transporter [DAT] (Catalogue No. MAB369) localization of DAT in a rat with a unilaterial 6-OH dopamine lesion. The transporter is se...
Anti-Dopamine Transporter, N-terminus, clone DAT-Nt
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, M, Mk, R | IC, IH, WB | Rat | Culture Supernatant | Monoclonal Antibody |
Description:
Anti-Dopamine Transporter, N-terminus, clone DAT-Nt
Trade Name:
Chemicon (Millipore)
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).
Molecular Weight:
~ 70-85 kDa
Epitope:
N-terminus
Immunogen:
N-terminus of human dopamine transporter fused to Glutathione S-transferase.
Clone:
DAT-Nt
Isotype:
IgG2aκ
Background Information:
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.
Species Reactivity:
- Human
- Mouse
- Monkey
- Rat
Application Notes:
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).
Control:
Positive Control: Brain (caudate, putamen, and nucleus accumbens) tissue
Purification Method:
Unpurified
Presentation:
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.
Storage Conditions:
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.
UniProt Number:
Entrez Gene Number:
Gene Symbol:
- SLC6A3
- DAT1
- ADHD
- ADD
- DAT
Alternate Names:
DAT
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.
Key Applications:
- Immunocytochemistry
- Immunohistochemistry
- Western Blotting
Entrez Gene Summary:
The 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]
UniProt Summary:
FUNCTION: 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.
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.
Brand Family:
Chemicon
Product Name:
Anti-Dopamine Transporter, N-terminus, clone DAT-Nt
Concentration:
Variable
Antibody Type:
Monoclonal Antibody
Qty/Pk:
100 μL
Format:
Culture Supernatant
Host:
Rat