Anti-Na+/H+ Exchanger 1 (NHE-1) (extracellular) Antibody

SLC9A1
    Cat #: ANX-010
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)RERSIGDVTTAPSE, corresponding to amino acid residues 54-67 of rat NHE-1 (Accession P26431). 1st extracellular loop.
    Accession (Uniprot) Number P26431
    Gene ID 24782
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - identical; human, pig, dog - 13/14 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, 0.05% 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 -20ºC.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ic, ifc, ih, lci, wb
    Western blot
    Western blot analysis of rat brain membranes (lanes 1 and 6), mouse brain lysate (lanes 2 and 7), human MCF-7 breast adenocarcinoma cells (lanes 3 and 8), human U-87 MG glioblastoma cells (lanes 4 and 9) and human THP-1 acute monocytic leukemia cells (lanes 5 and 10):
    1-5. Anti-Na+/H+ Exchanger 1 (NHE-1) (extracellular) Antibody (#ANX-010), (1:200).
    6-10. Anti-Na+/H+ Exchanger 1 (NHE-1) (extracellular) Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Expression of NHE-1 in mouse substantia nigra pars compacta
    Immunohistochemical staining of perfusion-fixed frozen mouse brain sections with Anti-Na+/H+ Exchanger 1 (NHE-1) (extracellular) Antibody (#ANX-010), (1:200), followed by goat-anti-rabbit-AlexaFluor-488. NHE-1 immunoreactivity (green) appears in neuronal profiles (arrows). Cell nuclei are stained with DAPI (blue).
    Indirect flow cytometry
    Indirect flow cytometry analysis of live intact THP-1 (human acute monocytic leukemia cells) cell line:
    ___ Cells + goat anti-rabbit-DyLight-488.
    ___ Cells + Anti-Na+/H+ Exchanger 1 (NHE-1) (extracellular) Antibody (#ANX-010), (1:20) + goat-anti-rabbit-DyLight-488.
    The control antigen is not suitable for this application.
    Live cell imaging / Immunocytochemistry
    Expression of NHE-1 in human MCF-7 cells
    Immunocytochemical staining of live intact human MCF-7 breast adenocarcinoma cells. A. Extracellular labeling of cells with Anti-Na+/H+ Exchanger 1 (NHE-1) (extracellular) Antibody (#ANX-010), (1:25), followed by goat anti-rabbit-AlexaFluor-594 secondary antibody (red). B. Live view of the cells. C. Merge of the two pictures.
    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. Orlowski, J. and Grinstein, S. (1997) J. Biol. Chem. 272, 22373.
    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-1 expression is ubiquitous and may serve as a housekeeping gene9.

    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
    Last update: 14/11/2018

    Anti-Na+/H+ Exchanger 1 (NHE-1) Antibody (#ANX-010) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot, immunohistochemistry, immunocytochemistry, live cell imaging, and indirect live cell flow cytometry applications. It has been designed to recognize NHE-1 from human, rat, and mouse samples.

    For research purposes only, not for human use