Guinea pig Anti-Kir4.1 (KCNJ10) Antibody

ATP-sensitive inward rectifier potassium channel 10, KAB-2, BIR10, BIRK1, Kir1.2, Potassium channel Inwardly rectifying subfamily J member 10
    Cat #: AGP-012
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Guinea pig
    Reactivity: h, m, r
    Immunogen
    Peptide (C)KLEESLREQAEKEGSALSVR, corresponding to amino acid residues 356-375 of rat Kir4.1 (Accession P49655). Intracellular, C-terminus.
    Accession (Uniprot) Number P49655
    Gene ID 29718
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Human, 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.
    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, ih, wb
    May also work in: ifc, ip
    Western blot
    Western blot analysis of rat brain membranes (lanes 1 and 4), mouse brain membranes (lanes 2 and 5) and human SH-SY5Y neuroblastoma cell lysates (lanes 3 and 6):
    1-3. Guinea pig Anti-Kir4.1 (KCNJ10) Antibody (#AGP-012), (1:800).
    4-6. Guinea pig Anti-Kir4.1 (KCNJ10) Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Expression of Kir4.1 in rat hippocampal dentate gyrus
    Immunohistochemical staining of rat frozen brain sections using Guinea pig Anti-Kir4.1 (KCNJ10) Antibody (#AGP-012), (1:300). A. Kir4.1 staining (green) reveals Kir4.1 clusters in the molecular layer (Mol). B. The same section was stained with mouse anti-glial fibrillary acidic protein (GFAP), (red) showing astrocyte profiles. C. Merge of Kir4.1 and GFAP images reveals colocalization on some branches of astrocytic fibers. In addition, Kir4.1 is expressed in the fine ramification of astrocytic fibers, thus creating a “halo-like” cloud around the GFAP astrocytic outline. DAPI (blue) counterstain reveals the outline of the dentate granule layer.
    Immunocytochemistry
    Expression of Kir4.1 in human U-87 MG glioblastoma cell line
    Immunocytochemical staining of fixed and permeabilized U-87 MG cells using Guinea pig Anti-Kir4.1 (KCNJ10) Antibody (#AGP-012), (1:100) followed by goat anti-guinea pig-AlexaFluor-488 secondary antibody (green). Cell nuclei were visualized using Hoechst 33342 (blue).
    References
    1. Takumi, T. et al. (1995) J. Biol. Chem. 270, 16339.
    2. Higashi, K. et al. (2001) Am. J. Physiol. 281, C922.
    3. Butt, A.M. et al. (2006) J. Cell Mol. Med. 10, 33.
    Scientific background

    Kir4.1 is a member of the family of inward rectifying K+ channels. The family includes 15 members that are structurally and functionally different from the voltage-dependent K+ channels.

    The family’s topology consists of two transmembrane domains that flank a single and highly conserved pore region with intracellular N- and C-termini. As is the case for the voltage-dependent K+ channels the functional unit for the Kir channels is composed of four subunits that can assemble as either homo or heteromers.

    Kir channels are characterized by a K+ efflux that is limited by depolarizing membrane potentials thus making them essential for controlling resting membrane potential and K+ homeostasis.

    Kir4.1 is a member of the Kir4 subfamily that includes one other member: Kir4.2. Kir4.1 can co-assemble with Kir4.2 but also with other Kir channels such as Kir2.1 and Kir5.1.

    The Kir4 subfamily has been classified as weak rectifiers with intermediate conductance. 

    Kir4.1, encoded by KCNJ10, is mainly expressed in brain, specifically in glia cells, but also in retina, ear and kidney1,2.

    It has been proposed that Kir4.1 has an essential role in glial K+ buffering, a process that re-uptakes the K+ released during neuronal activity into the intracellular interstitial space. Loss of Kir4.1 causes retinal defects and loss of endochoclear potential3.

    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:

    Guinea pig Anti-Kir4.1 (KCNJ10) Antibody
    Immuno-colocalization of Kir4.1 and α1-Syntrophin in rat fornix
    Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using Guinea pig Anti-Kir4.1 (KCNJ10) Antibody (#AGP-012), (1:400) and Anti-α1-Syntrophin Antibody (#APZ-021), (1:300). A. Kir4.1 staining (red) appears in blood vessel profiles (arrows) in the fornix. B. α1-Syntrophin staining in the same section (green) appears in several elements including blood vessels (arrows). C. Merge of the two images reveals partial co-localization of Kir4.1 and α1-Syntrophin in hilar cells. Cell nuclei are stained with DAPI (blue).

    Last update: 20/11/2018

    Guinea pig Anti-Kir4.1 (KCNJ10) Antibody (#AGP-012) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot, immunohistochemistry, and immunocytochemistry applications. It has been designed to recognize Kir4.1 potassium channel from human, rat, and mouse samples. The antigen used to immunize guinea pigs is the same as Anti-Kir4.1 (KCNJ10) Antibody (#APC-035) raised in rabbit. Our line of guinea pig antibodies enables more flexibility with our products such as immuno-colocalization studies, immunoprecipitation, etc.

    For research purposes only, not for human use
    Citations
    Immunohistochemistry citations
    1. Mouse brain sections. Also tested in Kir4.1-/- mice.
      Brasko, C. et al. (2016) Brain Struct. Funct. 222, 41.
    2. Mouse DRG sections (1:1000).
      Rajasekhar, P. et al. (2015) J. Biol. Chem. 290, 29051.
    Related Products
      • Antibodies for immuno-colocalization experiments
        1. Anti-Kir4.1 (KCNJ10)-ATTO-488 Antibody (#APC-035-AG). A fluorescent labeled primary antibody. It can be used in immuno-colocalization studies in conjunction with any of our antibodies raised in rabbit.