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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, M, Mk, R | ICC, IHC, WB | R | Culture Supernatant | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MAB369 |
| Brand Family | Chemicon® |
| Trade Name |
|
| Description | Anti-Dopamine Transporter Antibody, NT, clone DAT-Nt |
| Alternate Names |
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| 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 Parkinsons, Huntingtons and Chorea. |
| Product Information | |
|---|---|
| Format | Culture Supernatant |
| Control |
|
| 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. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This 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 |
|
| 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). |
| Biological Information | |
|---|---|
| Immunogen | N-terminus of human dopamine transporter fused to Glutathione S-transferase. |
| Epitope | N-terminus |
| Clone | DAT-Nt |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Rat |
| 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). |
| Isotype | IgG2aκ |
| Species Reactivity |
|
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| 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] |
| Gene Symbol |
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| Purification Method | Unpurified |
| UniProt Number |
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| 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. |
| Molecular Weight | ~ 70-85 kDa |
| Product Usage Statements | |
|---|---|
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| 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. |
| Packaging Information | |
|---|---|
| Material Size | 100 µL |