NKCC1 (D13A9) Rabbit mAb

8351:

Immunofluorescence , Immunoprecipitation , Western Blotting

NKCC1 (D13A9) Rabbit mAb recognizes endogenous levels of total NKCC1 protein. This antibody does not cross-react with NKCC2.

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg80 of human NKCC1 protein.

Western Blotting

Western blot analysis of extracts from HT-29, COLO 205, and DLD-1 cells using NKCC1 (D13A9) Rabbit mAb.

IF-IC

Confocal immunofluorescent analysis of MCF7 cells using NKCC1 (D13A9) Rabbit mAb (green). Blue pseudocolor= DRAQ5^® #4084 (fluorescent DNA dye).

The electroneutral cation-chloride-coupled co-transporter (SLC12) gene family comprises bumetanide-sensitive Na⁺ /K⁺ /Cl^- (NKCC), thiazide-sensitive Na⁺ /Cl^- , and K⁺ /Cl^- (KCC) co-transporters. SLC12A1/NKCC2 and SLC12A2/NKCC1 regulate cell volume and maintain cellular homeostasis in response to osmotic and oxidative stress (1). The broadly expressed NKCC1 is thought to play roles in fluid secretion (i.e. salivary gland function), salt balance (i.e. maintenance of renin and aldosterone levels), and neuronal development and signaling (2-7). During neuronal development, NKCC1 and KCC2 maintain a fine balance between chloride influx (NKCC1) and efflux (KCC2), which regulates γ-aminobutyric acid (GABA)-mediated neurotransmission (3). Increased NKCC1 expression in immature neurons maintains high intracellular chloride levels that result in inhibitory GABAergic signaling; KCC2 maintains low intracellular chloride levels and excitatory GABAergic responses in mature neurons (4,5,8). Deletion of NKCC1 impairs NGF-mediated neurite outgrowth in PC-12D cells while inhibition of NKCC1 with bumetanide inhibits re-growth of axotomized dorsal root ganglion cells (6,7). Defective chloride homeostasis in neurons is linked to seizure disorders that are ameliorated by butemanide treatment, indicating that abnormal NKCC1 and NKCC2 expression or signaling may play a role in neonatal and adult seizures (9-12). NKCC1 is found as a homodimer or within heterooligomers with other SLC12 family members. This transport protein associates with a number of oxidative- and osmotic-responsive kinases that bind, phosphorylate, and activate NKCC1 co-transporter activity (13-16). In response to decreased intracellular chloride concentrations, Ste20-related proline-alanine-rich kinase (SPAK) phosphorylates NKCC1 to increase co-transporter activity and promote chloride influx (16-19). Oxidative stress response kinase 1 (OSR1) also phosphorylates and activates NKCC1 in response to oxidative stress (14).

1. Hebert, S.C. et al. (2004) Pflugers Arch 447, 580-93.
2. Evans, R.L. et al. (2000) J Biol Chem 275, 26720-6.
3. Kim, S.M. et al. (2008) Am J Physiol Renal Physiol 295, F1230-8.
4. Khirug, S. et al. (2008) J Neurosci 28, 4635-9.
5. Kahle, K.T. et al. (2008) Nat Clin Pract Neurol 4, 490-503.
6. Nakajima, K. et al. (2007) Biochem Biophys Res Commun 359, 604-10.
7. Pieraut, S. et al. (2007) J Neurosci 27, 6751-9.
8. Ben-Ari, Y. (2002) Nat Rev Neurosci 3, 728-39.
9. Fukuda, A. (2005) Nat Med 11, 1153-4.
10. Dzhala, V.I. et al. (2005) Nat Med 11, 1205-13.
11. Jayakumar, A.R. et al. (2008) J Biol Chem 283, 33874-82.
12. Kahle, K.T. and Staley, K.J. (2008) Neurosurg Focus 25, E22.
13. Moore-Hoon, M.L. and Turner, R.J. (2000) Biochemistry 39, 3718-24.
14. Simard, C.F. et al. (2007) J Biol Chem 282, 18083-93.
15. Piechotta, K. et al. (2002) J Biol Chem 277, 50812-9.
16. Dowd, B.F. and Forbush, B. (2003) J Biol Chem 278, 27347-53.
17. Geng, Y. et al. (2009) J Biol Chem 284, 14020-8.
18. Smith, L. et al. (2008) J Biol Chem 283, 22147-56.
19. Gagnon, K.B. et al. (2006) Mol Cell Biol 26, 689-98.

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know !

• 4828 NKCC1 Antibody
• 3729 OSR1 Antibody
• 2281 SPAK Antibody
• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack

For Research Use Only. Not For Use In Diagnostic Procedures.