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NKCC1 (extracellular) Antibody

Affinity purified polyclonal antibody

  • WB - NKCC1 (extracellular) Antibody AG1162-025
     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 (extracellular) antibody (#AG1162), (1:200).
    4-6. Anti-NKCC1 (extracellular) antibody, preincubated with the control peptide antigen.
  • IHC - NKCC1 (extracellular) Antibody AG1162-025
    Expression of NKCC1 in rat cerebellum Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using Anti-NKCC1 (extracellular) antibody (#AG1162), (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.
  • ICC - NKCC1 (extracellular) Antibody AG1162-025
     Expression of NKCC1 in human U-87 MG cells  Immunocytochemical staining of live intact human U-87 MG glioblastoma cells. A. Extracellular staining of cells with Anti-NKCC1 (extracellular) antibody (#AG1162), (1:50), followed by goat anti-rabbit-AlexaFluor-594 secondary antibody (red). B. Live view of the cells. C. Merge of the two pictures.
Product Information
  • Applications Legend:
  • WB=Western Blot
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin-embedded Sections)
  • IHC-F=Immunohistochemistry (Frozen Sections)
  • IF=Immunofluorescence
  • FC=Flow Cytopmetry
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • IP=Immunoprecipitation
  • DB=Dot Blot
  • CHIP=Chromatin Immunoprecipitation
  • FA=Fluorescence Assay
  • IEM=Immunoelectronmicroscopy
  • EIA=Enzyme Immunoassay
Primary Accession P55011
Reactivity Human, Mouse, Rat
Host Rabbit
Clonality Polyclonal
Calculated MW 131447 Da
Homology Rat - identical; mouse - 13/14 amino acid residues identical.
Additional Information
Gene ID 6558
Other Names Solute carrier family 12 member 2, Basolateral Na-K-Cl symporter, Bumetanide-sensitive sodium-(potassium)-chloride cotransporter 1, SLC12A2, NKCC1
Related products for control experimentsControl peptide antigen (supplied with the antibody free of charge).
Target/Specificity Peptide (C)RRQAMKEMSIDQAK, corresponding to amino acid residues 828-841 of human NKCC1 (Accession P55011). 6th extracellular loop.
Dilution WB~~1:200-1:2000
Peptide Confirmation Confirmed by mass-spectrography and amino acid analysis.
Format Affinity purified antibody, lyophilized powder
Reconstitution 50 µl or 0.2 ml deionized water, depending on the sample size.
Antibody Concentration After Reconstitution 0.6 mg/ml.
Storage Before ReconstitutionLyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.
Storage After ReconstitutionThe reconstituted solution can be stored at 4ºC for up to 2 weeks. For longer periods, small aliquots should be stored at -20ºC or below. Avoid multiple freezing and thawing. The further dilutions should be made using a carrier protein such as BSA (1%). Centrifuge all antibody preparations before use (10000 × g 5 min).
Control Antigen Storage Before ReconstitutionLyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.
Control Antigen Reconstitution 100 µl DDW.
Control Antigen Storage After Reconstitution-20ºC.
Preadsorption Control 1 µg peptide per 1 µg antibody.
Formulation Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
Research Areas
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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 epithelia1. 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 protein2.   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 cells3. 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+-ATPase4. 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.


References 1. Haas M. (1994) Am. J. Physiol. 267, C869. 2. Delpire E. et al. (1999) Nat. Genet. 22, 192. 3. Haas M. and Forbush, B. 3rd (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.

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Cat# AG1162-025
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