Free shipping starts now, no minimum, no coupons required!

Anti-NKCC1 (SLC12A2) (extracellular) Antibody

Na+-K+-2Cl- Cotransporter 1, Basolateral Na-K-Cl symporter, Bumetanide-sensitive sodium-(potassium)-chloride cotransporter 1, BSC2, Solute carrier family 12 member 2

Cat #: ANT-071
Alternative Name Na+-K+-2Cl- Cotransporter 1, Basolateral Na-K-Cl symporter, Bumetanide-sensitive sodium-(potassium)-chloride cotransporter 1, BSC2, Solute carrier family 12 member 2
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide (C)RRQAMKEMSIDQAK, corresponding to amino acid residues 828-841 of human NKCC1 (Accession P55011). 6th extracellular loop.
Accession (Uniprot) Number P55011
Gene ID 6558
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Rat - identical; mouse - 13/14 amino acid residues identical.
RRID AB_2341018.
Purity Affinity purified on immobilized antigen.
Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
Isotype Rabbit IgG.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
Antibody concentration after reconstitution 0.6 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
Standard quality control of each lot Western blot analysis.
Applications: ic, if, ih, lci, wb
May also work in: ifc*, ip*
Western blot
  • Western blot analysis of rat brain membranes (lanes 1 and 4), mouse brain membranes (lanes 2 and 5) and rat lung lysate (lanes 3 and 6):
    Western blot analysis of rat brain membranes (lanes 1 and 4), mouse brain membranes (lanes 2 and 5) and rat lung lysate (lanes 3 and 6):
    1-3. Anti-NKCC1 (SLC12A2) (extracellular) Antibody (#ANT-071), (1:200).
    4-6. Anti-NKCC1 (SLC12A2) (extracellular) Antibody, preincubated with NKCC1/SLC12A2 (extracellular) Blocking Peptide (#BLP-NT071).
Immunohistochemistry
  • Expression of NKCC1 in rat cerebellum
    Expression of NKCC1 in rat cerebellum
    Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using Anti-NKCC1 (SLC12A2) (extracellular) Antibody (#ANT-071), (1:100). A. NKCC1 staining (red) is detected in the cerebellar molecular layer (Mol). B. DAPI counterstain (blue) reveals the outline of cerebellar layers. C. Merge of the two images.
Live cell imaging / Immunocytochemistry
  • Expression of NKCC1 in human U-87 MG cells
    Expression of NKCC1 in human U-87 MG cells
    Cell surface detection of NKCC1 in live intact human U-87 MG glioblastoma cells. A. Extracellular staining of cells with Anti-NKCC1 (SLC12A2) (extracellular) Antibody (#ANT-071), (1:50), followed by goat anti-rabbit-AlexaFluor-594 secondary antibody (red). B. Live view of the cells. C. Merge of the two pictures.
References
  1. Haas, M. (1994) Am. J. Physiol. 267, C869.
  2. Delpire E. et al. (1999) Nat. Genet. 22, 192.
  3. Haas, M. et al. (2000) Annu. Rev. Physiol. 62, 515.
  4. Ben-Ari, Y. et al. (2007) Physiol. Rev. 87, 1215.
  5. Delpire, E. et al. (2000) News. Physiol. Sci. 15, 309.
Scientific background

The Na-K-Cl cotransporters are a class of membrane proteins that transport Na+, K+, and Cl- ions into and out of cells in an electrically neutral manner, in most cases with a stoichiometry of 1Na+:1K+:2Cl-. Na-K-Cl cotransporters are present in a wide variety of cells and tissues, including reabsorptive and secretory epithelia, nerve and muscle cells, endothelial cells, fibroblasts, and blood cells.

Na-K-Cl cotransport plays a vital role in renal salt reabsorption and in salt secretion by intestinal, airway, salivary gland, and other secretory epithelia.1 Na-K-Cl cotransport activity is inhibited by "loop" diuretics, including the clinically efficacious agents, bumetanide and furosemide. The regulation of Na-K-Cl cotransport is mediated, at least in some cases, through direct phosphorylation of the cotransport protein.2

NKCC1 is one of two Na-K-Cl cotransporter isoforms that have been identified. NKCC1 is present in a wide variety of secretory epithelia and non-epithelial cells.3 NKCC1 is expressed in virtually all mammalian cells and is thought to play a housekeeping role in cell volume homeostasis and the common control of cytosolic ion content. NKCC1 does not use ATP but operates using electrochemical gradient for Na+ and K+ produced by Na+-K+-ATPase.4 There is considerable evidence that uptake of Cl in immature neurons is mediated by NKCC1. High expression of NKCC1 in immature neurons plays an important role in maintaining high intracellular Cl.5

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat
Last update: 25/07/2021

Anti-NKCC1 (SLC12A2) (extracellular) Antibody (#ANT-071) is a highly specific antibody directed against an epitope of the human protein. The antibody can be used in western blot, immunohistochemistry, live cell imaging, and immunocytochemistry applications. It has been designed to recognize NKCC1 from rat, mouse, and human samples.

For research purposes only, not for human use

Applications

Citations

Citations
Western blot citations
  1. Mouse brain lysate.
    Moriguchi, S. et al. (2018) Neuropharmacology 131, 291.
Immunohistochemistry citations
  1. Mouse eye sections.
    Frederikse, P.H. and Kasinathan, C. (2015) Mol. Vis. 21, 1142.

Specifications

Scientific Background

Shipping and Ordering information