Anti-KCNK2 (TREK-1) Antibody

Potassium channel subfamily K member 2, Outward rectifying potassium channel protein TREK-1, Twik-related K+ channel 1, K2P2.1, TPKC1
Cat #: APC-047
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide DPKSAAQNSKPRLSFSTK(C), corresponding to residues 8-25 of human KCNK2 (Accession O95069). Intracellular, N-terminus.
    Accession (Uniprot) Number O95069
    Gene ID 3776
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Rat - identical; mouse - 17/18 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, 0.025% 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:
    1. Anti-KCNK2 (TREK-1) Antibody (#APC-047), (1:200).
    2. Anti-KCNK2 (TREK-1) Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Rat hippocampus sections (Banerjee, A. et al. (2016) J. Neurochem. 138, 265.).
    Immunocytochemistry
    Mouse primary cortical astrocytes (1:100) (Hwang, E.M. et al. (2014) Nat. Commun. 5, 3227.).
    References
    1. Maingret, F. et al. (1999) J. Biol. Chem. 274, 26691.
    2. Lesage, F. and Lazdunski, M. (2000) Am. J. Physiol. Renal Physiol. 279, F793.
    3. Kim, D. (2003) Trends Pharmacol. Sci. 24, 648.
    4. Heurteaux, C. et al. (2006) Nat. Neurosci. 9, 1134.
    5. Woo, D.H. et al. (2012) Cell 151, 25.
    Scientific background

    KCNK2 (also named TWIK-related K+ channel, TREK-1 or K2P2.1) is a member of the 2-pore (2P) domain K+ channels family that includes at least 16 members. These channels show little time- or voltage-dependence and are considered to be “leak” or “background” K+ channels, thereby generating background currents which help set the membrane resting potential and cell excitation.

    The K2P channels have a signature topology that includes four transmembrane domains and two pore domains with intracellular N- and C- termini. 

    K2P channels are regulated by diverse physical and chemical stimuli including temperature, changes in intracellular pH, mechanical stretch, inhalation anesthetics, etc. The channels can then be subclassified based in their specific activators. KCNK2 can be integrated to a K2P subfamily that includes K2P4.1 (TRAAK) and K2P10.1 (TREK2) that are activated by intracellular unsaturated fatty acids such as arachidonic acid, lysophosphatidic acid and mechanical stretch. In addition, KCNK2 can also be activated by general anesthetics such as halothane and chloroform and intracellular acidification.

    KCNK2 expression in humans is largely restricted to the brain with some expression in ovary and small intestine while KCNK2 expression in rodents is more widespread.

    KCNK2 has an important role in mood regulation, as knockout mice show resistance to depression, suggesting that KCNK2 may be a potential target for anti-depressants.

    KCNK2 is involved in the fast release of glutamate from astrocytes. It requires the activation of Gαi, dissociation of Gβγ, followed by the opening of the glutamate permeable KCNK2 (TREK-1) K+ channel through its interaction with Gβγ. For this purpose, KCNK2 is localized at the cell surface of the cell body and processes of astrocytes.

    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-KCNK2 (TREK-1) Antibody
    Knockout validation of Anti-KCNK2 (TREK-1) Antibody in mouse hippocampus and expression of TREK-1 in mouse astrocytes. Immunohistochemical staining of mouse brain sections using Anti-KCNK2 (TREK-1) Antibody (#APC-047). A. TREK-1 staining (green) co-localizes with µ-opioid receptor (red) in astrocytes in CA1 region. B. TREK-1 expression (green) in mouse hippocampus (left panel). Lack of TREK-1 staining in TREK-1-/- mice confirms antibody specificity (right panel).Adapted from Woo, D.H. et al. (2018) Front. Cell. Neurosci. 12, 319. with permission of Frontiers.

    Last update: 25/03/2019

    Anti-KCNK2 (TREK-1) Antibody (#APC-047) is a highly specific antibody directed against an epitope of the human protein. The antibody can be used in western blot, immunohistochemistry, and immunocytochemistry applications. It has been designed to recognize TREK-1 from mouse, rat, and human samples.

    For research purposes only, not for human use
    Citations
    Published figures using this product
    Expression of TREK-1 in rat hippocampus.
    Immunohistochemical staining of rat brain sections using Anti-KCNK2 (TREK-1) Antibody (#APC-047). TREK-1 staining (red) in the hippocampus is detected in the stratum radiatum, in the neuropil. Similar diffused staining is observed for GLAST. GFAP staining, the astrocyte marker is also shown.
    Adapted from Zhou, M. et al. (2009) J. Neurosci. 29, 8551. with permission of the Society for Neuroscience.
    KO validation citations
    1. Immunohistochemical staining of mouse brain sections. Also tested in TREK-1-/- mice.
      Woo, D.H. et al. (2018) Front. Cell. Neurosci. 12, 319.
    2. Western blot analysis of mouse spinal cord lysate. Also tested in TREK-1-/- mice.
      Fang, Y. et al. (2017) J. Neurochem. 141, 236.
    Western blot citations
    1. Mouse spinal cord lysate. Also tested in TREK-1-/- mice.
      Fang, Y. et al. (2017) J. Neurochem. 141, 236.
    2. Rat astrocyte lysate (1:100).
      Banerjee, A. et al. (2016) J. Neurochem. 138, 265.
    3. Rat astrocyte lysate.
      Wu, X. et al. (2013) J. Mol. Neurosci. 49, 499.
    4. Mouse myometrium lysate.
      Monaghan, K. et al. (2011) J. Physiol. 589, 1221.
    5. Human adrenocortical carcinoma H295R cells.
      Brenner, T. and O'Shaughnessy, K.M. (2008) Am. J. Physiol. 295, E1480.
    Immunohistochemistry citations
    1. Mouse brain sections. Also tested in TREK-1-/- mice.
      Woo, D.H. et al. (2018) Front. Cell. Neurosci. 12, 319.
    2. Mouse brain sections.
      Cai, Y. et al. (2017) Neurobiol. Learn. Mem. 145, 199.
    3. Mouse spinal cord sections.
      Fang, Y. et al. (2017) J. Neurochem. 141, 236.
    4. Rat Hippocampus sections.
      Banerjee, A. et al. (2016) J. Neurochem. 138, 265.
    5. Mouse myometrium sections.
      Monaghan, K. et al. (2011) J. Physiol. 589, 1221.
    6. Rat brain sections.
      Zhou, M. et al. (2009) J. Neurosci. 29, 8551.
    Immunocytochemistry citations
    1. Rat astrocytes.
      Banerjee, A. et al. (2016) J. Neurochem. 138, 265.
    2. Mouse primary cortical astrocytes (1:100).
      Hwang, E.M. et al. (2014) Nat. Commun. 5, 3227.
    3. Rat astrocyte culture.
      Wu, X. et al. (2013) J. Mol. Neurosci. 49, 499.
    More product citations
    1. Du, Y. et al. (2016) Front. Cell. Neurosci. 10, 13.
    2. de la Pena, E. et al. (2012) PLoS ONE 7, e52475.
    3. Woo, D.H. et al. (2012) Cell 151, 25.
    4. Lembrechts, R. et al. (2011) Histochem. Cell. Biol. 136, 371.
    5. Baker, S.A. et al. (2010) J. Urol. 183, 793.
    6. Mazella, J. et al. (2010) PLoS Biol. 8, e1000355.
    7. Zhou, M. et al. (2009) J. Neurosci. 29, 8551.
    8. Brenner, A. et al. (2008) Am. J. Physiol. Endocrinol. Metab. 295, E1480.
    9. Voloshyna, I. et al. (2008) Cancer Res. 68, 1197.
    10. Magra, M. et al. (2007) Am. J. Physiol. Cell Physiol. 292, C1053.
    11. Zhao, M. et al. (2007) J. Neurosci. 27, 12025.
    12. Gardener, M.J. et al. (2004) Br. J. Pharmacol. 142, 192.
    13. Bai, X. et al. (2005) Reproduction 129, 525.
    14. Murbartian, J. et al. (2005) J. Biol. Chem. 280, 30175.
    15. Eaton, M.J. et al. (2004) NeuroReport 15, 321.
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