Anti-Synaptophysin Antibody

Major synaptic vesicle protein p38, SYP
    Cat #: ANR-013
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: m, r
    May also work in: h
    Immunogen
    Peptide CRQTGNT(S)KELRD, corresponding to amino acid residues 178-190 of rat synaptophysin (Accession P07825). 2nd intravesicular loop.
    Accession (Uniprot) Number P07825
    Gene ID 24804
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Rat, mouse, human - 12/13 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 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: ih, wb
    May also work in: ic, ifc, ip
    Western blot
    Western blot analysis of rat brain membranes (lanes 1 and 4), mouse brain membranes (lanes 2 and 5) and rat PC12 pheochromocytoma cell line lysate (lanes 3 and 6):
    1-3. Anti-Synaptophysin Antibody (#ANR-013), (1:400).
    4-6. Anti-Synaptophysin Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Expression of Synaptophysin in rat DRG
    Immunohistochemical staining of rat dorsal root ganglia (DRG) frozen sections using Anti-Synaptophysin Antibody (#ANR-013), (1:100). Synaptophysin (red) is expressed in DRG neurons. Hoechst 33342 (blue) shows nuclear staining and is used as the counterstain.
    Colocalization of NaV1.8 and Synaptophysin in rat DRG
    Immunohistochemical staining of rat DRG frozen section using Anti-NaV1.8 (SCN10A)-ATTO-594 Antibody (#ASC-016-AR) and Anti-Synaptophysin Antibody (#ANR-013). A. NaV1.8 staining (red). B. Synaptophysin staining (green). C. Merged image demonstrates a partial overlap in the distribution of Nav1.8 and Synaptophysin within the DRGs. DAPI is used as the counterstain (blue).
    References
    1. Jahn, R. et al. (1985) Proc. Natl. Acad. Sci. U.S.A. 82, 4137.
    2. Navone, F. et al. (1986) J. Cell Biol. 103, 2511.
    3. Wiedenmann, B. and Franke, W.W. (1985) Cell 41, 1017.
    4. Johnston, P.A. et al. (1989) J. Biol. Chem. 264, 1268.
    5. Kwon, S.E. and Chapman, E.R. (2012) J. Biol. Chem. 287, 35658.
    6. Kwon, S.E. and Chapman, E.R. (2011) Neuron 70, 847.
    7. Schmitt, V. et al. (2009) Neuroscience 162, 234.
    8. Tarpey, P.S. et al. (2009) Nat. Genet. 41, 535.
    Scientific background

    Synaptophysin is the major integral membrane protein of small synaptic vesicles1-3. It belongs to a family of proteins which includes synaptogyrin and synaptoporin. The protein contains 4 transmembrane domains and cytoplasmic N- and C-termini4. There are two intravesicular loops that contain disulfide bonds. The protein is N-glycosylated on the first intravesicular loop, a modification important for the synaptic vesicle targeting of synaptophysin. Indeed, a mutant form of the protein, unable to undergo glycolysation, leads to the accumulation of the protein in the cell body of neurons indicating that N-glycosylation is required for the synaptic localization of synaptophysin5. The C-terminal tail is extensively phosphorylated on Tyr residues. However, the biological significance of the post-translational modification is not yet determined6.

    Although exact and specific role of synaptophysin remains a mystery, a study suggests that the protein is required for efficient endocytosis of synaptic vesicles in cultured hippocampal neurons6.

    Since synaptophysin is exclusively detected in synaptic vesicles, it is widely used as a marker for presynaptic vesicles.

    Genetic screening in humans and behavioral studies in rodents show that loss of synaptophysin expression or expression of a truncated form of the protein may be attributed to mental retardation and/or learning deficits7,8. Interestingly though, synaptic transmission in synaptophysin knockout mice remains normal6.

    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: 30/10/2018

    Anti-Synaptophysin Antibody (#ANR-013) is a highly specific antibody directed against an epitope of the rat protein. It can be used in western blot and immunohistochemistry applications. It has been designed to recognize synaptophysin from mouse, rat, and human samples.

    For research purposes only, not for human use
    Citations
    Western blot citations
    1. Mouse brain lysate.
      Yang, P. et al. (2016) Exp. Neurol. 279, 178.
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