NHE3/SLC9A3 Antibody (19F5)

Product Discontinued

NHE3/SLC9A3 Antibody (19F5) Summary

• Immunogen: Fusion protein representing the C-terminal 131 amino acids of rabbit NHE3. [UniProt# P26432]
• Localization: Membrane; multi-pass membrane.
• Specificity: This antibody is specific for NHE3. It does not show any cross-reactivity with any other NHE isoforms.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: SLC9A3
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • ELISA
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Frozen 1:10-1:500
  • Immunohistochemistry-Paraffin 1:10-1:500
  • Immunoprecipitation 1:10-1:500
  • Western Blot 0.5 ug/ml
• Application Notes: In Western blot, a band can be seen at ~93 kDa.
• Theoretical MW: 93 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reactivity Notes

Immunogen sequence has 83% homology to mouse. Does not react with human.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: Tris-Glycine and 0.15M NaCl
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein G purified

Alternate Names for NHE3/SLC9A3 Antibody (19F5)

• isoform 3
• MGC126718
• NHE3
• NHE3MGC126720
• SLC9A3
• solute carrier family 9 (sodium/hydrogen exchanger), member 3
• Solute carrier family 9 member 3

Background

Na+/H+ exchanger (NHE) family members are integral membrane proteins which play key role in catalyzing the extrusion of intracellular proton (H+) ions in exchange for extracellular sodium (Na+) ions for regulation of cellular pH as well as transepithelial ion/water transport. NHE3 or SLC9A3 is one of the best characterized NHEs that is a major proton extruding system driven by inward Na+ ion chemical gradient and is implicated in pH regulation to eliminate acids generated during metabolism or to tackle any potential adverse conditions in tissue microenvironment. NHE3 exists as a multi-pass membrane protein in cell membranes and is expressed most abundantly in colon and small intestine, followed by kidney and stomach, as well as in ventrolateral region of the medulla oblongata. In intestinal epithelial cells, it localizes to the ileal brush border and SGK1 mediated phosphorylation at Ser-661 has been shown to be associated with its enhanced cell membrane localization. NHE3 binds to SLC9A3R1 and SLC9A3R2, and interacts with CHP1, CHP2, SHANK2 PDZD3 and (interactions may decrease in response to elevated calcium ion levels). NHE3 is regulated by several hormones, neurotransmitters, and associated signaling systems such as cAMP, cGMP, and elevated intracellular calcium via mechanisms that are largely partially understood. NHE3 knock-out mice suffers from chronic diarrhea and altered salt as well as water homeostasis.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for NHE3/SLC9A3 Antibody (NB110-61586)(10)

Publications using NB110-61586

• Man A, Chen M, Wu Z et al. Renal Effects of Fetal Reprogramming With Pentaerythritol Tetranitrate in Spontaneously Hypertensive Rats Front Pharmacol. 2020-04-29 [PMID: 32410988] (WB, Rat)
• Saad S, Zhang J, Yong R et al. Role of the EGF receptor in PPARgamma-mediated sodium and water transport in human proximal tubule cells. Diabetologia 2013-01-31 [PMID: 23370527] (IHC-P, Rat)
• Kocinsky HS, Girardi AC, Biemesderfer D, Nguyen T, Mentone S, Orlowski J, Aronson PS. Use of phospho-specific antibodies to determine the phosphorylation of endogenous Na+/H+ exchanger NHE3 at PKA consensus sites. Am J Physiol Renal Physiol;289(2):F249-58. 2005-08-01 [PMID: 15687252]
• Sabolic I, Herak-Kramberger CM, Ljubojevic M, Biemesderfer D, Brown D. NHE3 and NHERF are targeted to the basolateral membrane in proximal tubules of colchicine-treated rats. Kidney Int;61(4):1351-64. 2002-04-01 [PMID: 11918742] (WB, ICC/IF, Rat)
• Bagnis C, Marsolais M, Biemesderfer D, Laprade R, Breton S. Na+/H+-exchange activity and immunolocalization of NHE3 in rat epididymis. Am J Physiol Renal Physiol;280(3):F426-36. 2001-03-01 [PMID: 11181404] (IF/IHC, Rat)
• Girardi AC, Titan SM, Malnic G, Reboucas NA. Chronic effect of parathyroid hormone on NHE3 expression in rat renal proximal tubules. Kidney Int;58(4):1623-31. 2000-10-01 [PMID: 11012896] (WB, IF/IHC, Rat)
• Biemesderfer D, Nagy T, DeGray B, Aronson PS. Specific association of megalin and the Na+/H+ exchanger isoform NHE3 in the proximal tubule. J Biol Chem;274(25):17518-24. 1999-06-18 [PMID: 10364184] (IP, Rabbit)
• Zhang Y, Norian JM, Magyar CE, Holstein-Rathlou NH, Mircheff AK, McDonough AA. In vivo PTH provokes apical NHE3 and NaPi2 redistribution and Na-K-ATPase inhibition. Am J Physiol;276(5 Pt 2):F711-9. 1999-05-01 [PMID: 10330053] (WB, Rat)
• Biemesderfer D, DeGray B, Aronson PS. Membrane topology of NHE3. Epitopes within the carboxyl-terminal hydrophilic domain are exoplasmic. J Biol Chem. 273(20):12391-6. 1998-05-15 [PMID: 9575193] (ELISA, IP, WB, Rabbit)
• Biemesderfer, D et al. Monoclonal antibodies for high-resolution localization of NHE3 in adult and neonatal rat kidney. Am J Physiol;273(2 Pt 2):F289-99. 1997-08-01 [PMID: 9277590] (WB, ICC/IF, ELISA, IHC-Fr, Rabbit, Rat)

FAQs for NHE3/SLC9A3 Antibody (NB110-61586). (Showing 1 - 1 of 1 FAQs).

1. The molecular weight of NHE3 is 92kDa but I see a band above 100 kDa.  I am confused, because the size of these protein should not be greater than 100KD. Can you please help explain?  
   • Higher than predicted molecular weight seen on WB is not uncommon if the protein is posttranslationally modified, especially glycosylated, and such is the case for NHE3. Please note that NHE3 gets phosphorylated and glycosylated (Uniprot ID P48764) and potentially, these modifications are adding ~10kDa extra to the predicted molecular weight. If you want to confirm it 100%, you could try de-glycosylating your protein and then performing a WB on it. 

Bioinformatics

• Gene Symbol: SLC9A3