NKCC1 (Sodium, Potassium, Chloride Cotransporter 1, BSC2, CCC1)

The electroneutral cation-chloride-coupled co-transporter (SLC12) gene family is comprised of bumetanide-sensitive Na+/K+/Cl- (NKCC), thiazide-sensitve 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 g-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). \n\nNKCC1 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). \n\nApplications:\nSuitable for use in Western Blot and Immunoprecipitation. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 1:1000. Incubate membrane with diluted antibody in 5% w/v BSA, 1X TBS, 0.1% \n Tween-20 at 4 degrees C with gentle shaking, overnight. \nImmunoprecipitation: 1:50.\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

N2800-01B

Size

100ul

Applications

IP, WB

Hosts

Rabbit

Reactivities

Hum

Form

Supplied in 10mM sodium HEPES pH 7.5, 150mM NaCl, 0.1mg/ml BSA, 50% glycerol.

P Type

Pab

Purity

Purified by Protein A affinity chromatography.

Isotype

IgG

References

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.

Additional Info

Recognizes human NKCC1.