Anti-Na+/H+ Exchanger 2 (NHE-2) Antibody

Sodium/hydrogen exchanger 2, SLC9A2
    Cat #: ANX-002
    Alternative Name Sodium/hydrogen exchanger 2, SLC9A2
  • Lyophilized Powder
  • Antigen Incl.
  • Type: Polyclonal
    Host: Rabbit
    Reactivity: h, m, r
    Immunogen
    • Peptide (C)RASEPGNRKGRLGNEK, corresponding to amino acid residues 797-812 of rat NHE-2 (Accession P48763). Intracellular, C-terminus.
    • Anti-Na+/H+ Exchanger 2 (NHE-2) Antibody
    Accession (Uniprot) Number P48763
    Gene ID 24783
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - identical; human - 12/16 amino acid residues identical.
    RRID AB_11219158.
    Purity Affinity purified on immobilized antigen.
    Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% 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).
    Negative 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.
    Negative control antigen reconstitution 100 µl double distilled water (DDW).
    Negative 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: if, ih, wb
    May also work in: ic*, ifc*, ip*
    Western blot
    • Anti-Na+/H+ Exchanger 2 (NHE-2) Antibody
      Western blot analysis of rat kidney membranes (lanes 1 and 4), mouse kidney lysates (lanes 2 and 5) and rat stomach lysates (lanes 3 and 6):
      1-3. Anti-Na+/H+ Exchanger 2 (NHE-2) Antibody (#ANX-002), (1:200).
      4-6. Anti-Na+/H+ Exchanger 2 (NHE-2) Antibody, preincubated with the negative control antigen.
    • Anti-Na+/H+ Exchanger 2 (NHE-2) Antibody
      Western blot analysis of human Colo-205 colorectal carcinoma cell lysates:
      1. Anti-Na+/H+ Exchanger 2 (NHE-2) Antibody (#ANX-002), (1:200).
      2. Anti-Na+/H+ Exchanger 2 (NHE-2) Antibody, preincubated with the negative control antigen.
    Immunohistochemistry
    • Anti-Na+/H+ Exchanger 2 (NHE-2) Antibody
      Expression of Na+/H+ exchanger 2 in rat colon
      Immunohistochemical staining of rat colon sections (paraffin-embedded) using Anti-Na+/H+ Exchanger 2 (NHE-2) Antibody (#ANX-002), (1:50). A. NHE-2 staining is shown in red. B. Cell nuclei were labeled with DAPI (blue). C. Merge of the two images.
    References
    1. Counillon, L. and Pouyssegur, J. (2000) J. Biol. Chem. 275, 1.
    2. Kemp, G. et al. (2008) Channels 2, 329.
    3. Counillon, L. et al. (1994) Biochemistry 33, 10463.
    4. Tse, C. et al. (1994) Biochemistry 33, 12954.
    5. Paris, S. and Poouyssegur, J. (1983) J. Biol. Chem. 258, 3503.
    6. Aronson, P.S. et al. (1983) J. Biol. Chem. 258, 6767.
    7. Yu, F.H. et al. (1993) J. Biol. Chem. 268, 25536.
    8. Aronson, P.S. et al. (1982) Nature 299, 161.
    9. Bookstein, C. et al. (1997) Am. J. Physiol. 273, 1496.
    10. Chambrey, R. et al. (1998) Am. J. Physiol. 275, 379.
    11. Schultheis, P.J. et al. (1998) Nat. Genet. 19, 282.
    12. Malakooti, J. et al. (1999) Am. J. Physiol. 277, 383.
    Scientific background

    In order to function in optimal conditions, cells must maintain a close to neutral intracellular pH. They have adopted various mechanisms in order to do so, one of which is via Na+/H+ exchangers (NHEs). Genes belonging to this group are expressed along a very broad range of organisms and are essential for protecting cells against intracellular acidification1.

    To date, nine genes have been identified in mammals; NHE1-9. These membrane proteins have 10-12 transmembrane domains depending on whether a splice variant is expressed and an intracellular N-terminal. The C-terminal domain can be either intracellular or extracellular, also depending whether a splice variant of the protein is involved. The C-terminal part of the protein also undergoes posttranslational modification such as phosphorylation2. Both NHE-1 and NHE-2 have an extracellular loop which is glycosylated1,3,4.

    Under physiological conditions, the Na+/H+ exchanger mediates the exchange of one extracellular Na+ ion for one intracellular proton, thereby keeping the overall charge neutral1. The extracellular binding site of Na+ is not selective as it can also bind Li+ and H+ 1,5,6.  K+ ions inhibit NHE-1 but have no effect on NHE-27. The activation of NHE-1 and NHE-2 is sensitive to intracellular acidic pH. Under physiological conditions, both exchangers are not active and upon a drop of intracellular pH, they are rapidly activated1,5,8.

    NHE-2 is detected in the intestine, kidney and parietal cells2,9-11. It is also detected in skeletal muscle and testis2,12.

    Application key:

    CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, 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: 24/01/2020

    Anti-Na+/H+ Exchanger 2 (NHE-2) Antibody (#ANX-002) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize NHE-2 from human, rat, and mouse samples.

    For research purposes only, not for human use

    Applications

    Specifications

    Scientific Background

    Citations

    Citations
    Western blot citations
    1. Human Barrett's esophageal cell lysate (1:200).
      Laczko, D. et al. (2016) Am. J. Physiol. 311, G16.
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