Anti-Noradrenaline Transporter (NET) (extracellular) Antibody

Sodium-dependent noradrenaline transporter, Norepinephrine transporter, SLC6A2
    Cat #: AMT-002
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: m, r
    May also work in: h
    Immunogen
    Peptide (C)KLLNASVLGDHTKYSK, corresponding to amino acid residues 189-204 of mouse Noradrenaline Transporter (Accession O55192). 2nd extracellular loop.
    Accession (Uniprot) Number O55192
    Gene ID 20538
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Rat - identical; human - 14/16 amino acid residues identical.
    Purity Affinity purified on immobilized antigen.
    Formulation Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
    Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
    Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
    Antibody concentration after reconstitution 0.8 mg/ml.
    Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
    Control antigen storage before reconstitution Lyophilized powder can be stored intact at room temperature for 2 weeks. For longer periods, it should be stored at -20°C.
    Control antigen reconstitution 100 µl double distilled water (DDW).
    Control antigen storage after reconstitution -20ºC.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ic, ih, lci, wb
    May also work in: ifc, ip
    Western blot
    Western blot analysis of rat (lanes 1 and 3) and mouse (lanes 2 and 4) brain membranes:
    1, 2. Anti-Noradrenaline Transporter (NET) (extracellular) Antibody (#AMT-002), (1:400).
    3, 4. Anti-Noradrenaline Transporter (NET) (extracellular) Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Expression of Noradrenaline Transporter (NET) in rat hippocampal dentate gyrus
    Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using Anti-Noradrenaline Transporter (NET) (extracellular) Antibody (#AMT-002), (1:200). A. NET (green) is visualized in axonal processes (arrows) in the hilus. B. The relation to layers was explored using mouse anti-GAP43 (red). There are less axonal processes in the outer molecular layer (asterisk) compared to the inner molecular layer (arrows). C. Overlay of A and B shows co-localization of NET and GAP43 in the outer molecular layer.
    Live cell imaging / Immunocytochemistry
    Expression of Noradrenaline Transporter (NET) in live intact rat PC12 cells
    Immunocytochemical staining of live intact rat pheochromocytoma PC12 cells. Cells were stained with Anti-Noradrenaline Transporter (NET) (extracellular) Antibody (#AMT-002), (1:100), followed by goat anti-rabbit-AlexaFluor-594 secondary antibody (red). Cell nuclei (blue) were visualized using cell-permeable Hoechst 33342.
    References
    1. Ramamoorthy, S. et al. (2010) Pharmacol. Ther. 129, 220.
    2. Siuta, M.A. et al. (2010) PLoS Biol. 8, e1000393.
    3. Madras, B.K. et al. (2005) Biol. Psychiatry 57, 1397.
    4. Gainetdinov, R.R. and Caron, M.G. (2003) Annu. Rev. Pharmacol. Toxicol. 43, 261.
    5. Moron, J.A. et al. (2002) J. Neurosci. 22, 389.
    6. Greengard, P. (2001) Science. 294, 1024.
    7. Kitayama, S. et al. (2001) Neurosci. Lett. 312, 108.
    8. Miller, G.W. et al. (1999) Trends. Pharmacol. Sci. 20, 424.
    9. Giros, B. et al. (1993) Trends. Pharmacol. Sci. 14, 43.
    10. Carlsson, A. (1987) Annu. Rev. Neurosci. 10, 19.
    Scientific background

    Many physiological, endocrine and behavioral functions are determined and regulated by monoamine signaling1,2. Many brain disorders such as depression, drug abuse, schizophrenia, attention deficit hyperactivity disorder (ADHD) are caused by the malfunction of monoaminergic transmission1-3.  The intensity of monoaminergic signaling is determined by the availability of the monoamine, which is in turn determined in part by its uptake from the extracellular milieu via monoamine transporters. These transporters include DAT, SERT, and NET, responsible for uptaking dopamine, serotonin and noradrenaline respectively, and recycling them back for release3-5.

    While the activity of each transporter is faithful to its neurotransmitter, NET has been shown to clear dopamine in DAT deprived or low DAT regions such as the brain cortex6-8.

    DAT, SERT and NET are members of the Na+/Cl- dependent membrane transporter family which also includes other members. These transporters consist of 12 transmembrane domains and intracellular N- and C-termini. NET also has a significant extracellular loop between transmembrane regions three and four, which contains various glycosylation sites9. Like its counterparts, NET’s intracellular N- and C-terminal domains are also subject to phosphorylation and protein-protein interactions important for modulating its activity and localization. In addition alternative splicing has also been shown to regulate NET’s expression and function10.

    NET is specifically expressed on noradrenaline nerve terminals, and is also expressed in the periphery, such as adrenal glands and placenta. NET malfunction is largely associated with attention and mood, as well as various cardiovascular disorders. NET is also a target for psychostimulants such as cocaine and amphetamines which disrupt its function, thereby causing an increase of noradrenaline in synaptic clefts3,9.

    Application key:

    CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

    Species reactivity key:

    H- Human, M- Mouse, R- Rat
    Image & Title:

    Immuno-colocalization of Noradrenaline Transporter (NET) and Neurokinin Receptor 1 (NK1) in rat brain stem.Immunohistochemical staining of perfusion-fixed frozen rat brain sections using Anti-Noradrenaline Transporter (NET) (extracellular) Antibody (#AMT-002), (1:400) and Anti-Neurokinin Receptor 1 (NK1) (extracellular)-ATTO-488 Antibody (#ATR-001-AG), (1:80). A. NET staining (red) in section of rat locus coeruleus. B. NK1 staining (green) in same section. C. Merge of the two images reveals several cells expressing both NET and NK1 (arrows). Cell nuclei are stained with DAPI (blue).

    Last update: 01/11/2018

    Alomone Labs is pleased to offer a highly specific antibody directed against an epitope of mouse noradrenaline transporter. Anti-Noradrenaline Transporter (NET) (extracellular) Antibody (#AMT-002) can be used in western blot immunohistochemistry and immunocytochemistry applications. It has been designed to recognize NET from human, mouse and rat samples.

    For research purposes only, not for human use