Anti-KCNJ5 (Kir3.4) Antibody

GIRK4, G protein-activated inward rectifier potassium channel 4, Cardiac inward rectifier, CIR, KATP-1
Cat #: APC-027
Alternative Name GIRK4, G protein-activated inward rectifier potassium channel 4, Cardiac inward rectifier, CIR, KATP-1
  • KO Validated
  • Lyophilized Powder
  • Antigen Incl.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
      • Peptide RNAMNQDMEIGVT(C), corresponding to amino acid residues 6-18 of rat Kir3.4 (Accession P48548). Intracellular, N-terminus.
    Accession (Uniprot) Number P48548
    Gene ID 29713
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse, human, pig - identical.
    Purity Affinity purified on immobilized antigen.
    Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.025% NaN3.
    Isotype Rabbit IgG.
    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 analysis of rat heart membranes:
        1. Anti-KCNJ5 (Kir3.4) Antibody (#APC-027), (1:200).
        2. Anti-KCNJ5 (Kir3.4) Antibody, preincubated with the control antigen.
      • Rat heart sections (1:20) (Atkinson, A.J. et al. (2013) J. Am. Heart Assoc. 2, e000246.).
      • Mouse HL-1 cells (1:500) (Nobles, M. et al. (2010) Pflugers Arch. 460, 99.).
      • Kir3.4 is a member of the G-protein regulated inward-rectifier K+ (GIRK) channel subfamily which is part of an inward-rectifier K+ channel superfamily. The GIRK subfamily comprises four members in mammals (Kir3.1- Kir3.4) that present the common topology of the inward-rectifier superfamily: two transmembrane domains flanking a highly conserved pore region with the N and C-terminus located intracellularly.

        Kir3.4 and the other Kir3 family subunits can be activated by neurotransmitters and other factors via the activation of G-protein coupled receptors. Binding of the corresponding ligand to the G-protein receptor induces the dissociation of G α-GTP from the Gβγ dimer. The latter directly binds to Kir3 and activates the channel. 

        Kir3.4 expression is largely confined to the heart, where it co-assembles with Kir3.1 to form the prototypical muscarinic-gated K+ channel KACh. Indeed, knockout mice for Kir3.4 showed impaired heart rate following vagal nerve stimulation.

        A peptide toxin originating from the Apis mellifera bee venom, Tertiapin (#STT-250) was shown to be a potent blocker of Kir3.4 containing channels (8.6 nM for the Kir3.1/3.4 combination).4

    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-KCNJ5 (Kir3.4) Antibody
    Knockout validation of Anti-KCNJ5 (Kir3.4) Antibody in mouse heart.
    Immunohistochemical staining of mice heart using Anti-KCNJ5 (Kir3.4) Antibody (#APC-027). A. GIRK4 staining in GIRK4 KO mice atrial sections. B. GIRK4 staining in GIRK4 KO mice ventricular sections. C. GIRK4 staining in GIRK1 KO mice atrial sections and (D) ventricular sections. Staining demonstrates that the channel is localized to the outer sarcolemma and the ventricular t-tubular system. Scale bar = 10 μm.
    Adapted from Liang, B. et al. (2014) with permission of the European Society of Cardiology.
    Last update: 10/06/2019

    Alomone Labs is pleased to offer a highly specific antibody directed against an epitope of the rat Kir3.4 channel. Anti-KCNJ5 (Kir3.4) Antibody (#APC-027) can be used in western blot and immunohistochemistry applications. It has been designed to recognize Kir3.4 from human, rat and mouse samples.

    For research purposes only, not for human use
    Citations
      • Immunohistochemical staining of mouse, rat and human heart sections (1:100). Also tested in GIRK4-/- mice.
        Liang, B. et al. (2014) Cardiovasc. Res. 101, 175.
      • Mouse brain lysate (1:500).
        Goodfellow, N.M. et al. (2014) J. Neurosci. 34, 6107.
      • Mouse, rat and human heart sections (1:100). Also tested in mouse GIRK4-/-.
        Liang, B. et al. (2014) Cardiovasc. Res. 101, 175.
      • Rat heart sections (1:20).
        Atkinson, A.J. et al. (2013) J. Am. Heart Assoc. 2, e000246.
      • Mouse HL-1 cells (1:500).
        Nobles, M. et al. (2010) Pflugers Arch. 460, 99.
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