Application

Research CategoryNeuroscience

Anti-phospho-NHE3 (Ser552), clone 14D5, Cat. No. MABN2415, is a highly specific mouse monoclonal antibody that targets Sodium/hydrogen exchanger 3 and has been tested for use in ELISA, Immunofluorescence, and Western Blotting.

Immunofluorescence Analysis: A representative lot detected phospho-NHE3 (Ser552) in Immunofluorescence applications (Kocinsky, H.S., et. al. (2005). Am J Physiol Renal Physiol. 289(2):F249-58).

Western Blotting Analysis: A representative lot detected phospho-NHE3 (Ser552) in Western Blotting applications (Kocinsky, H.S., et. al. (2005). Am J Physiol Renal Physiol. 289(2):F249-58; Kocinsky, H.S., et. al. (2007). Am J Physiol Renal Physiol. 293(1):F212-8).

ELISA Analysis: A representative lot detected phospho-NHE3 (Ser552) in ELISA applications (Kocinsky, H.S., et. al. (2005). Am J Physiol Renal Physiol. 289(2):F249-58).

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Sodium/hydrogen exchanger 3 (UniProt: P26433; also known as Na(+)/H(+) exchanger 3, NHE-3, Solute carrier family 9 member 3) is encoded by the Slc9a3 (Nhe3) gene (Gene ID: 24784) in rat. NHE-3 is a multi-pass membrane protein that is involved in pH regulation to eliminate acids generated by active metabolism or to counter adverse environmental conditions. It is a major proton extruding system driven by the inward sodium ion chemical gradient. NHE-3 is shown to be most abundant in colon and small intestine and is also present in stomach and in proximal tubes and outer medulla of kidney. In kidneys it mediates the majority of sodium bicarbonate reabsorption by the proximal tubules. NHE-3 can be phosphorylated and regulated by multiple kinases, including PKA, SGK1, and CK2 and its phosphorylation is critical in its regulation. Parathyroid hormone is shown to enhance PKA-mediated phosphorylation of Serine 552, however, this phosphorylation does not affect the activity of NHE-3. (Ref.: Kocinsky, HS., et al. (2007). Am J Physiol Renal Physiol 293 (1); F212-F218).

Immunogen

KLH-conjugated linear peptide corresponding to 22 amino acids surrounding phospho Serine 552 from the C-terminal half of rat Sodium/hydrogen exchanger 3 (NHE3).

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Protein G purified

Format: Purified

Purified mouse monoclonal antibody IgG2a in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Quality

Evaluated by Western Blotting in COS-7 cells either transfected with wild type NHE3, mutated NHE3 (S552A), or untransfected COS-7 (WT) cell lysates.

Western Blotting Analysis: 2 µg/mL of this antibody detected Sodium/hydrogen exchanger 3 in lysate from COS-7 cells transfected with wild- type NHE-3 (pSer552). It did not detect NHE-3 in lysate from untransfected COS-7 cells or those transfected with mutated NHE-3 (S552A).

Specificity

This mouse monoclonal antibody (clone 14D5) specifically detects Sodium/hydrogen exchanger 3 phosphorylated on Serine 552.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Target description

~85 kDa observed; 93.11 kDa calculated. Uncharacterized bands may be observed in some lysate(s).