Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody

IKCa1, IK1, Intermediate conductance Ca2+-activated K+ channel protein 4, Gardos channel
    Cat #: ALM-051
  • Lyophilized Powder
  • Shipped at Room Temp.
  • Type: Monoclonal
    Source: Mouse
    Reactivity: h, m, r
    Immunogen
    Synthetic peptide mapping to the 3rd extracellular loop of human KCNN4 (Accession O15554).
    Accession (Uniprot) Number O15554
    Gene ID 3783
    Purity Affinity purified from cultured hybridoma medium.
    Formulation Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
    Isotype IgM.
    Clone 6C1.
    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 or 100 µl double distilled water (DDW), depending on the sample size.
    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).
    Standard quality control of each lot Western blot analysis.
    Applications: ic, ifc, ih, ip, lci, wb
    Western blot
    Western blot analysis using Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody (#ALM-051), (1:250):
    1. HEK cells transfected with control vector.
    2. HEK cells transfected with human KCa3.1.
    Western blot analysis using Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody (#ALM-051), (1:250):
    1. Mouse MS1 endothelial cells.
    2. Rat IEC-6 intestinal epithelial cells.
    3. Human LN-CaP prostate carcinoma cells.
    4. Human THP-1 acute monocytic leukemia cells.
    Immunohistochemistry
    Immuno-colocalization of KCa3.1 (SK4) and TRPC1 in mouse brain
    Immunohistochemical staining of mouse brain sections using Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody (#ALM-051) and Anti-TRPC1 Antibody (#ACC-010). TRPC1 staining (blue) is detected in neuroblasts and outside neuroblasts as well (in astrocytes). KCa3.1 (red) is detected in neuroblasts. Merged image demonstrates the partial co-localization between the two.
    Adapted from Turner, K.L. et al. (2014) Cereb. Cortex. 24, 2388. with permission of Oxford University Press.
    Indirect flow cytometry
    Indirect flow cytometry analysis of live intact Raji (human Burkitt's lymphoma B cells) cell line:
    ___ Cells + goat-anti-mouse-Cy5.
    ___ Cells + mouse IgM isotype control + goat-anti-mouse-Cy5.
    ___ Cells + Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody (#ALM-051), (1:20) + goat-anti-mouse-Cy5.
    Indirect flow cytometry analysis of live intact THP-1 (human acute monocytic leukemia cells) cell line:
    ___ Cells + Goat-anti-mouse-Cy5.
    ___ Cells + Mouse IgM isotype control+ Goat-anti-mouse-Cy5.
    ___ Cells + Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody (#ALM-051), (1:20) + Goat-anti-mouse-Cy5.
    Live cell imaging / Immunocytochemistry
    Expression of KCNN4 in live intact human LN-CaP prostate carcinoma cells
    Immunocytochemical staining of live intact human LN-CaP prostate carcinoma cells with Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody (#ALM-051) (1:20), followed by goat-anti-mouse-DyLight-594 secondary antibody (red) (A). B. Live view of the cells. C. Merge of the two images.
    References
    1. Ghanshani, S. et al. (2000) J. Biol. Chem. 275, 37137.
    2. Hoffman, J.F. et al. (2003) Proc. Natl. Acad. Sci. U.S.A. 100, 7366.
    Scientific background

    KCNN4 (KCa3.1, SK4) is a member of the Ca2+ activated K+ channel family that shares the characteristic of being activated by intracellular Ca2+. The channel has an intermediate conductance, is voltage insensitive and is activated by Ca2+ in the submicromolar range. The channel has a similar topology to that of KV channels, that is, six transmembrane domains and intracellular N- and C-termini. 

    KCNN4 is widely expressed in epithelial, endothelial and cells of hematopoietic origin. In erythrocytes (red blood cells) it has been identified as the molecular correlate of the so-called Gardos channel.  

    The functional role of the channel is to set the cell membrane potential at negative values so as to aid in the electrochemical transport of other ions such as Cl- and Ca2+. Indeed, KCNN4 has a key role in sustaining the Ca2+ influx in activated T lymphocytes and in regulating Cl- secretion from colon epithelium. Therefore, specific blockers of the KCNN4 channel have been proposed for the treatment of several diseases including autoimmune diseases, secretory diarrhea and sickle cell anemia.

    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:

    Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) AntibodyImmuno-colocalization of KCa3.1 (SK4) and TRPC1 in mouse brainImmunohistochemical staining of mouse brain sections using Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody (#ALM-051) and Anti-TRPC1 Antibody (#ACC-010). YFP denotes neuroblasts. TRPC1 staining (blue) is detected in neuroblasts and outside neuroblasts as well (in astrocytes). KCa3.1 (red) is detected in neuroblasts. Merged image of all three staining (lower right panel) demonstrates the partial co-localization between the three.Adapted from Turner, K.L. and Sontheimer, H. (2013) Cereb. Cortex 24, 2388. with permission of Oxford University Press.

    Last update: 09/12/2018

    Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody (#ALM-051) is a highly specific monoclonal antibody directed against an epitope of the human channel. The antibody can be used in western blot, immunocytochemistry, immunohistochemistry, and indirect flow cytometry applications. It has been designed to recognize KCNN4 from human, rat, and mouse samples.

    For research purposes only, not for human use
    Citations
    Published figures using this product
    Immuno-colocalization of TRPV4 and KCa3.1 in human BSM cells
    Immunocytochemical staining of human bronchial smooth muscle (HBSM) cells using Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody (#ALM-051), (green) and Anti-TRPV4 Antibody (#ACC-034), (red).
    Adapted from Yu, Z. et al. (2017) Front. Pharmacol. 8, 559. with permission of Frontiers.
    Immuno-colocalization of KCa3.1 and P2Y2 in human ovarian carcinoma
    Immunohistochemical staining of human ovarian carcinoma biopsies using Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody (#ALM-051) and rabbit Anti-P2Y2 Receptor Antibody (#APR-010). Positive co-expression is detected in ovarian neoplastic cells (merged image). Expression of KCa3.1 is also observed in stromal cells (*); however, P2Y2 does not co-localize with KCa3.1 in these cells.
    Adapted from Robles-Martinez, L. et al. (2017) Sci. Rep. 7, 4340. with permission of Nature Publishing Group.
    Expression of KCNN4 channel in human cytotrophoblasts
    Immunocytochemical staining of human cytotrophoblast using Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody (#ALM-051). A. Methanol fixed cells (15 hrs). B. Methanol fixed cells (66 hrs).
    Adapted from Diaz, P. et al. (2014) with kind permission of Dr. Diaz, P. and Dr. Greenwood, S.L. from Maternal and Fetal Health Research Centre, Institute of Human Development University of Manchester, Manchester, United Kingdom and St. Mary’s Hospital, Central Manchester University Hospitals National Health Service (NHS) Foundation Trust, Manchester Academic Health Science Centre, Manchester, United Kingdom.
    SK4 (KCNN4) channel is expressed in hESC-CMs
    Immunocytochemical staining of hESC-CMs. Cells were stained with Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody (#ALM-051) followed by anti-mouse-Dylight405 secondary antibody (green). α-Actinin staining was used as a control (red). Upper panels are 18 days old beating clusters. Lower panels are dissociated hESC-CMs.
    Adapted from Weisbrod, D. et al. (2013) Proc. Natl. Acad. Sci. U.S.A. 110, E1685. with permission of the National Academy of Sciences.
    Colocalization and expression of KCa3.1, ClC-3 and B2R in glioma cells.
    A-C. Immunocytochemical staining of human D54 glioma cells using Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody (#ALM-051). KCa3.1, ClC-3 and B2R co-localize to the edges of cells. D. Western blot analysis of a number of glioma cells using Anti-CLC-3 (CLCN3) Antibody (#ACL-001) and Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody.
    Adapted from Cuddapah, V.A. et al. (2013) with permission of the Society for Neuroscience.
    Western blot citations
    1. Human ovarian cancer SKOV-3 cell lysates.
      Robles-Martinez, L. et al. (2017) Sci. Rep. 7, 4340.
    2. Human dendritic cells.
      Crottes, D. et al. (2016) Cell Calcium 59, 198.
    3. Mouse mesenteric arteries lysate.
      Yap, F.C. et al. (2016) Am. J. Physiol. 310, H1151.
    4. Mouse heart lysate (1:600).
      Ju, C.H. et al. (2015) Cell. Physiol. Biochem. 36, 1305.
    Immunoprecipitation citations
    1. Mouse CD4+ T-cells.
      Srivastava, S. et al. (2016) eLife 19, 16093.
    Immunohistochemistry citations
    1. Human ovarian tumor sections.
      Robles-Martinez, L. et al. (2017) Sci. Rep. 7, 4340.
    2. Mouse brain sections.
      Turner, K.L. et al. (2014) Cereb. Cortex. 24, 2388.
    Immunocytochemistry citations
    1. Human ovarian cancer SKOV-3 cells.
      Robles-Martinez, L. et al. (2017) Sci. Rep. 7, 4340.
    2. Human bronchial smooth muscle (HBSM) cells.
      Yu, Z. et al. (2017) Front. Pharmacol. 8, 559.
    3. Human dendritic cells.
      Crottes, D. et al. (2016) Cell Calcium 59, 198.
    4. Mouse CCDcl1 cells (1:100).
      Li, Y. et al. (2016) PLoS ONE 11, e0155006.
    5. Human cytotrophoblasts.
      Diaz, P. et al. (2014) PLoS ONE 9, e90961.
    More product citations
    1. Yi, M. et al. (2017) J. Neuroinflammation 14, 203.
    2. Cuddapah, V.A. et al. (2013) J. Neurosci. 33, 1427.
    3. Weisbrod, D. et al. (2013) Proc. Natl. Acad. Sci. U.S.A. 110, E1685.
    4. Bagher, P. et al. (2012) Proc. Natl. Acad. Sci. U.S.A. 109, 18174.
    Related Products
      • Products for control experiments
        1. Anti-KCNN4 (KCa3.1, SK4) Antibody (#APC-064). This antibody recognizes a different epitope and thus can also be used as a control for detecting KCNN4.