Cell Signaling Technology

Product Pathways - Neuroscience

NKCC1 (D2O8R) XP® Rabbit mAb (Rodent Specific; IF Specific) #85403

No. Size Price
85403S 100 µl ( 200 tests ) ¥3,580.00 现货查询 购买询价
85403 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
IF-F 1:200 Mouse,Rat, Endogenous 160-200 Rabbit IgG

Species cross-reactivity is determined by western blot.

Applications Key: IF-F=Immunofluorescence (Frozen),

Specificity / Sensitivity

NKCC1 (D2O8R) XP® Rabbit mAb (Rodent specific; IF specific) recognizes endogenous levels of total NKCC1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of mouse NKCC1 protein.

IF-F

IF-F

Confocal immunofluorescent analysis of mouse small intestine (positive, left) or liver (negative, right), using NKCC1 (D208R) XP® Rabbit mAb (Rodent Specific; IF specific) (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

IF-F

IF-F

Confocal immunofluorescent analysis of rat cerebellum (positive, left) and liver (negative, right) using

NKCC1 (D208R) XP® Rabbit mAb (Rodent Specific; IF specific) (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Background

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.

Application References

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

DRAQ5 is a registered trademark of Biostatus Limited.

DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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