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Home › Products › Transporters, Exchangers and Pumps › Na+/H+ Exchangers › Antibodies

Certificate of Analysis

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

Sodium/hydrogen exchanger 1, SLC9A1, APNH

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Overview

Cat #: ANX-010
Alternative Name Sodium/hydrogen exchanger 1, SLC9A1, APNH
Lyophilized Powder yes
Type: Polyclonal
Host: 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.
RRID AB_2341021.
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).
Standard quality control of each lot Western blot analysis.
Applications: ic, if, ifc, ih, lci, wb
May also work in: ip*
Western blot
  • Human MCF-7 breast adenocarcinoma cells, U-87 MG glioblastoma cells and THP-1 acute monocytic leukemia cells. Rat and mouse brain membranes (1:200).
  • 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):
    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 Na+/H+ Exchanger 1/NHE-1 (extracellular) Blocking Peptide (#BLP-NX010).
Immunohistochemistry
  • Mouse brain sections (1:200).
Indirect flow cytometry
  • Human THP-1 acute monocytic leukemia cells (1:20).
  • The blocking peptide is not suitable for this application.
Live cell imaging / Immunocytochemistry
  • Human live and intact MCF-7 breast adenocarcinoma cells (1:25).
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, 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
Lyophilized Powder
For research purposes only, not for human use
Last Update: 10/04/2023

Specifications

Citations

Citations

Applications

Scientific Background

Specific Control Product

  • Na+/H+ Exchanger 1/NHE-1 (extracellular) Blocking Peptide (#BLP-NX010) is the original antigen used for immunization during Anti-Na+/H+ Exchanger 1 (NHE-1) (extracellular) Antibody (#ANX-010) generation. The blocking peptide binds and ‘blocks’ Anti-Na+/H+ Exchanger 1/NHE-1 (extracellular) primary antibody, this makes it a good negative reagent control to help confirm antibody specificity in western blot and immunohistochemistry applications. This control is also often called a pre-adsorption control.

    Na+/H+ Exchanger 1/NHE-1 (extracellular) Blocking Peptide (#BLP-NX010)

Related Products

Antibodies

  1. Anti-Na+/H+ Exchanger 2 (NHE-2) Antibody (#ANX-002)
  2. Anti-Na+/H+ Exchanger 3 (NHE-3) Antibody (#ANX-033)
  3. Anti-Na+/H+ Exchanger 5 (NHE-5) Antibody (#ANX-005)
  4. Anti-Na+/H+ Exchanger 6 (NHE-6) Antibody (#ANX-006)

Pharmacological tools

Blockers/Antagonists: small molecules
  1. Amiloride hydrochloride (#A-140)
  2. 5-(N,N-Dimethyl)amiloride hydrochloride (#D-165)
  3. HMA (#H-130)

Explorer kits & Research packs

Explorer kits
  1. Na+/H+ Exchanger (NHE) Antibody Explorer Kit (#AK-610)

General Protocols

  • Blocking Peptides – Controls for better results
  • Blocking Peptide Protocol for Western Blot (WB)
  • Sample Preparation for Cell Lines
  • Immunohistochemistry (IHC) Protocols for Frozen Sections: Indirect Methods
  • Flow Cytometry Protocols for Live Cells: Indirect and Direct Methods
  • Sample Preparation Protocols for Tissues
  • Western Blot (WB) Protocol

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