Anti-Vesicular Acetylcholine Transporter (VAChT) Antibody

Solute carrier family 18 member 3, SLC18A3
    Cat #: ACT-003
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: m, r
    Immunogen
    Peptide (C)DAVRLREVQGKDGGE, corresponding to amino acid residues 495-509 of rat VAChT (Accession Q62666). Cytoplasmic, C-terminus.
    Accession (Uniprot) Number Q62666
    Gene ID 60422
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - 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.
    Specificity Not recommended for human samples.
    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 2 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ih, wb
    May also work in: ic, ifc, ip
    Western blot
    Western blot analysis of mouse (lanes 1 and 3) and rat (lanes 2 and 4) brain lysates:
    1,2. Anti-Vesicular Acetylcholine Transporter (VAChT) Antibody (#ACT-003), (1:200).
    3,4. Anti-Vesicular Acetylcholine Transporter (VAChT) Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Expression of VAChT in rat medial septum and spinal cord
    Immunohistochemical staining of perfusion-fixed frozen rat brain sections using Anti-Vesicular Acetylcholine Transporter (VAChT) Antibody (#ACT-003), (1:400), followed by goat-anti-rabbit-AlexaFluor-488. A. VAChT staining (green) in rat medial septum is detected in neuronal soma (arrows). B. Staining of spinal cord sections reveals VAChT labeling in neuronal soma (arrows). Cell nuclei are stained with DAPI (blue).
    References
    1. Ribeiro, F.M. et al. (2006) J. Neurochem. 97, 1.
    2. Okuda, T. and Haga, T. (2003) Neurochem. Res. 28, 483.
    3. Bravo, D.T. et al. (2004) J. Neurochem. 91, 766.
    4. de Castro, B.M. et al. (2009) Mol. Cell. Biol. 29, 5238.
    5. Nguyen, M.L. et al. (2000) Biochemistry 37, 13400.
    6. Parsons, S.M. (2000) FASEB J. 14, 2423.
    7. Masson, J. et al. (1999) Pharmacol. Rev. 51, 439.
    Scientific background

    In cholinergic neurons, acetylcholine is synthesized from choline and acetyl-coenzyme A (acetyl-CoA). Vesicular acetylcholine transporter (VAChT) transports acetylcholine from the cytoplasm to the lumen of synaptic vesicles where following depolarization, acetylcholine is released in the synaptic cleft and activates muscarinic and nicotinic receptors located on postsynaptic membranes1-3. Acetylcholine released in the synaptic cleft is rapidly hydrolyzed into choline and acetate. Since choline is not de novo synthesized and thereby only made available through diet uptake, choline is recycled back into the cells in order to regenerate acetylcholine1,2.

    VAChT is a member of the SLC18 family of transporters. It has 12 membrane spanning domains, with extracellular N- and C-terminal tails. The transporter exchanges cytoplasmic acetylcholine for two vesicular protons4-6.

    VAChT is a selective marker of cholinergic neurons and could be used therefore to determine the loss of cholinergic neurons in Alzheimer’s or Alzheimer’s related disease7. VAChT knockout mice die after birth. They display a lack of stimulated release of acetylcholine and an underdeveloped neuromuscular junction4.

    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:

    Anti-Vesicular Acetylcholine Transporter (VAChT) AntibodyImmuno-colocalization of Vesicular Acetylcholine Transporter and p75NTR in rat medial septum.Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using rabbit Anti-Vesicular Acetylcholine Transporter (VAChT) Antibody (#ACT-003), (1:200) and Mouse Anti-Rat p75 NGF Receptor (extracellular) Antibody (#AN-170), (1:200). A. VAChT staining (green) appears in several neuronal cells. B. p75NTR (red) also stains neuronal cells. C. Merge of the two images shows colocalization of VAChT and p75NTR in some cells (horizontal arrows), while other cells express only VAChT (vertical arrows). Cell nuclei are stained with DAPI (blue).

    Last update: 27/11/2018

    Anti-Vesicular Acetylcholine Transporter (VAChT) Antibody (#ACT-003) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize VAChT from mouse and rat samples. The antibody is not suited to recognize VAChT from human samples.

    For research purposes only, not for human use