Anti-KV1.3 (KCNA3) (extracellular)-Biotin Antibody

Potassium voltage-gated channel subfamily A member 3
    Cat #: APC-101-B
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide KDYPASTSQDSFEA(C), corresponding to amino acid residues 263-276 of human KV1.3 (Accession number P22001). Extracellular loop between domains S1 and S2.
    Accession (Uniprot) Number P22001
    Gene ID 3738
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Rat, mouse - 12/14 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.
    Label Biotin.
    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 double distilled water (DDW).
    Antibody concentration after reconstitution 1 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: fc, ih, wb
    May also work in: ic, ip, lci
    Western blot
    Western blot analysis of human Jurkat T cell leukemia cell lysate (lanes 1 and 3) and mouse brain lysate (lanes 2 and 4):
    1-2. Anti-KV1.3 (KCNA3) (extracellular)-Biotin Antibody (#APC-101-B), (1:1000).
    3-4. Anti-KV1.3 (KCNA3) (extracellular)-Biotin Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Immuno-colocalization of KV1.3 and KV1.5 in mouse cerebellum
    Immunohistochemical staining of mouse perfusion-fixed frozen brain sections using Anti-KV1.3 (KCNA3) (extracellular)-Biotin Antibody (#APC-101-B) (1:400), and Anti-KV1.5 (KCNA5)-ATTO-550 Antibody (#APC-004-AO), (1:60). A. KV1.3 staining (green) is detected in Bergmann glia soma and processes in the molecular layer (Mol) (arrow). B. Same section shows staining for KV1.5 (red). C. Merge of the two images suggests considerable co-localization in the soma of Bergmann glia. In the molecular layer, the distribution of KV1.5 is diffuse, unlike the discrete staining for KV1.3 in glial processes. Cell nuclei were visualized with DAPI (blue).
    Direct flow cytometry
    Direct flow cytometry analysis of live intact human Jurkat T cell leukemia cell line:
    ___ Cells + Streptavidin-Alexa647.
    ___ Cells + Anti-KV1.3 (KCNA3) (extracellular)-Biotin Antibody (#APC-101-B), (1:10) + Streptavidin-Alexa647.
    The control antigen is not suitable for this application.
    References
    1. Chandy, K.G. et al. (2001) Toxicon 39, 1269.
    2. Koo, G.C. et al. (1997) J. Immunol. 158, 5120.
    Scientific background

    KV1.3 belongs to the Shaker family of voltage-dependent K+ channels. The channel, encoded by KCNA3, is widely expressed in the brain, lung and osteoclasts and in several cell populations of hematopoietic origin. It is in these cells (particularly T lymphocytes) that KV1.3 function has centered a lot of attention. It was found that KV1.3 is the main channel responsible for maintaining the resting potential in quiescent cells and regulating the Ca2+ signaling that is indispensable for normal T lymphocyte activation.

    Based on the central role of KV1.3 in regulating the initiation of an immune response, the channel has been recognized as a potential target for immunosuppressant drugs. The central role of KV1.3 in immune system cells created a real need for a specific antibody that would be able to work in flow cytometry applications.

    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: 27/11/2018

    Anti-KV1.3 (KCNA3) (extracellular) Antibody (#APC-101) is a highly specific antibody directed against an extracellular epitope of the human protein. The antibody can be used in western blot, immunohistochemistry, live cell imaging, and indirect flow cytometry applications. It has been designed to recognize KV1.3 potassium channel from human, mouse, and rat samples.

    Anti-KV1.3 (KCNA3) (extracellular)-Biotin Antibody (#APC-101-B) is directly labeled with biotin. Strepavidin tagged with HRP or with a fluorescent probe can then be used to detect the protein. The biotin/strepavidin system is ideal for minimizing cross-reactivity when same species antibodies are simultaneously used. Anti-KV1.3 (KCNA3) (extracellular)-Biotin Antibody has been tested in western blot, immunohistochemistry and direct flow cytometry and is specially suited to experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use
    Citations
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        1. Guinea pig Anti-KV1.3 (KCNA3) (extracellular) Antibody (#AGP-005) is raised in guinea pig and can be used in immuno-colocalization studies in conjunction with any of our antibodies raised in rabbit. This antibody has been raised against the same epitope as #APC-101.