Anti-Serotonin Transporter (SERT) (extracellular) Antibody

Sodium-dependent serotonin transporter, 5-HT transporter, 5-HTT, 5-HTTLPR, SLC6A4
    Cat #: AMT-004
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)EMRNEDVSEVAKD, corresponding to amino acids residues 388-400 of rat SERT (Accession P31652). 4th extracellular loop.
    Accession (Uniprot) Number P31652
    Gene ID 25553
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse, human - 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 -20°C.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ic, ifc, ih, lci, wb
    Western blot
    Western blot analysis of mouse (lanes 1 and 3) and rat (lanes 2 and 4) brain lysates:
    1-2. Anti-Serotonin Transporter (SERT) (extracellular) Antibody (#AMT-004), (1:200).
    3-4. Anti-Serotonin Transporter (SERT) (extracellular) Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Expression of SERT in mouse brain
    Immunohistochemical staining of immersion-fixed, free floating mouse brain frozen sections using Anti-Serotonin Transporter (SERT) (extracellular) Antibody (#AMT-004), (1:400). A. SERT (green) was visualized in axonal fibers of the hippocampal dentate gyrus. B. Axonal fibers of neurons expressing gamma amino butyric acid (GABA) were labeled with mouse anti parvalbumin (red). C. Merge of the two images demonstrates separate axonal processes (no co-localization).
    Expression of SERT in rat brain
    Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using Anti-Serotonin Transporter (SERT) (extracellular) Antibody (#AMT-004), (1:400). A. SERT (green) was visualized in axonal fibers of the rat hippocampal CA1 region. B. Axonal fibers of neurons expressing gamma amino butyric acid (GABA) were labeled with mouse anti parvalbumin (red). C. Merge of the two images demonstrates separate axonal processes (no co-localization).
    Indirect flow cytometry
    Indirect flow cytometry analysis in live intact human T cell leukemia (Jurkat) cell line:
    ___ Unstained cells + goat-anti-rabbit-Cy5.
    ___ Cells + Anti-Serotonin Transporter (SERT) (extracellular) Antibody (#AMT-004), (1:25) + goat-anti-rabbit-Cy5.
    The control antigen is not suitable for this application.
    Live cell imaging / Immunocytochemistry
    Expression of serotonin transporter in rat PC12 cells
    Immunocytochemical staining of live intact rat pheochromocytoma PC12 cells. A. Cells were stained with Anti-Serotonin Transporter (SERT) (extracellular) Antibody (#AMT-004), (1:100), followed by goat anti-rabbit-AlexaFluor-594 secondary antibody (red). B. Cell nuclei were visualized using Hoechst 33342 (blue). C. Live view of the cells.
    References
    1. Haenisch, B. and Bönisch, H. (2011) Pharmacol. Ther. 129, 352.
    2. Ramamoorthy, S. et al. (2010) Pharmacol. Ther. 129, 220.
    3. Kim, D.K. et al. (2005) Neuropharmacology 49, 798.
    4. Mathews, T.A. et al. (2004) J. Neurosci. Meth. 140, 169.
    5. Gainetdinov, R.R. and Caron, M.G. (2003) Annu. Rev. Pharmacol. Toxicol. 43, 261.
    6. Greengard, P. (2001) Science. 294, 1024.
    7. Miller, G.W. et al. (1999) Trends. Pharmacol. Sci. 20, 424.
    8. Amara, S.G. and Kuhar, M.J. (1993) Annu. Rev. Neurosci. 16, 73.
    9. 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 (5-HT) and noradrenaline respectively, and recycling them back for release3-5.

    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. Like its counterparts, SERT’s intracellular N- and C-terminal domains are also subject to phosphorylation and protein-protein interactions important for modulating its activity and localization6.

    SERTs are evidently expressed in serotonergic neurons, and are also expressed in various peripheral tissues including specialized cells of the gut, placenta, lung, blood lymphocytes and platelets6.

    SERT knockout mice are viable and as a consequence exhibit increased extracellular levels of 5-HT and significantly decreased levels of brain tissue 5-HT due to deficient recycling and re-accumulation of serotonin by SERT7,8. Phenotypes displayed by SERT knockout mice include stress sensitivity, obesity and various behavioral changes9

    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
    Last update: 21/11/2018

    Anti-Serotonin Transporter (SERT) (extracellular) Antibody (#AMT-004) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot, immunohistochemistry, immunocytochemistry, indirect flow cytometry, and live cell imaging applications. It has been designed to recognize SERT from human, mouse, and rat samples.

    For research purposes only, not for human use
    Citations
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