NKCC1 (D13A9) Rabbit mAb

• 产品编号：CST-8351S      品牌：CST       原厂货号：8351S
• 产品分类：抗体 > 一抗 > 蛋白特异性一抗
• 应用分类：

描述：

NKCC1 (D13A9) Rabbit mAb 兔单抗识别内源性的NKCC1总蛋白。此抗体与磷酸化的NKCC2蛋白没有交叉反应。该单克隆抗体由合成的人类NKCC1蛋白Arg80位点附近肽段免疫动物而制成。在电中性阳离子-氯耦合共转运（SLC12）基因家族包括布美他尼敏感的Na+/H+/Cl-（NKCC），噻嗪类敏感的Na+/Cl-，K+/CL-（KCC）共转运体。SLC12A1/NKCC2和SLC12A2/NKCC1调节细胞体积和维持细胞响应渗透和氧化应激时的稳态（1）。广泛表达NKCC1被认为在液体分泌（如唾液腺功能），水盐平衡（如维持肾素和醛固酮水平），以及神经元的发育和信号传导中发挥作用（2-7）。在神经发育中，NKCC1和KCC2在在氯离子涌入（NKCC1）和流出（KCC2）中维持良好平衡，调节γ-氨基丁酸（GABA）介导的神经传导（3）。在未成熟神经元中增加的NKCC1表达可以维持细胞内的氯的高水平，产生抑制性GABA能信号；KCC2在成熟神经元中保持细胞内氯的低水平，产生兴奋性GABA能反应（4,5,8）。删除NKCC1会损害神经生长因子介导的PC-12D细胞的突起生长，用布美他尼抑制NKCC1会抑制切断的背根神经节细胞的重新生长（6,7）。氯稳态缺陷的神经元与癫痫相关，这种癫痫可被butemanide治疗改善，这表明NKCC1和NKCC2异常表达或其信号通路可能在新生儿和成人癫痫中发挥作用（9-12）。NKCC1是作为二聚体和与其它SLC12家族成员形成异寡聚体被发现的。这种运输蛋白与氧化反应和渗透反应激酶结合，结合、磷酸化和激活NKCC1共转运体的活性（13-16）。为了应对降低的细胞内氯离子浓度，STE20相关的富含脯氨酸丙氨酸的蛋白激酶（SPAK）磷酸化NKCC1，增加共转运活动，促进氯离子涌入（16-19）。氧化应激反应激酶1（OSR1）在氧化应激反应中磷酸化和激活NKCC1（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.

原厂资料：

Homology

Species predicted to react based on 100% sequence homology: Bovine, Pig

Specificity / Sensitivity

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

Source / Purification

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

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.

注意事项：

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150
mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02%
sodium azide. Store at –20°C. Do not aliquot the antibody.