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Anti-NKCC2 (SLC12A1) (extracellular) Antibody

Na+-K+-2Cl- Cotransporter 2, Kidney-specific Na-K-Cl symporter, Bumetanide-sensitive sodium-(potassium)-chloride cotransporter 2, BSC1, Solute carrier family 12 member 1

Cat #: ANT-072
Alternative Name Na+-K+-2Cl- Cotransporter 2, Kidney-specific Na-K-Cl symporter, Bumetanide-sensitive sodium-(potassium)-chloride cotransporter 2, BSC1, Solute carrier family 12 member 1
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: m, r
May also work in: h*
Immunogen
  • Peptide (C)GYKKNWRKAPLSELE, corresponding to amino acid residues 776-790 of rat NKCC2 (Accession P55016). 6th extracellular loop.
Accession (Uniprot) Number P55016
Gene ID 25065
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Mouse - identical; human - 13/15 amino acid residues identical.
RRID AB_2756718.
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 0.8 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: wb
May also work in: ic*, ifc*, ih*, ip*, lci*
Western blot
  • Western blot analysis of rat (lanes 1 and 3) and mouse (lanes 2 and 4) kidney membranes:
    Western blot analysis of rat (lanes 1 and 3) and mouse (lanes 2 and 4) kidney membranes:
    1,2. Anti-NKCC2 (SLC12A1) (extracellular) Antibody (#ANT-072), (1:400).
    3,4. Anti-NKCC2 (SLC12A1) (extracellular) Antibody, preincubated with NKCC2/SLC12A1 (extracellular) Blocking Peptide (#BLP-NT072).
References
  1. Ares, G.R. et al. (2011) Am. J. Physiol. 301, 1143.
  2. Gamba, G. and Friedman, P.A. (2009) Pflugers Arch. 458, 61.
Scientific background

Na+-K+-2Cl cotransporter (NKCC2) belongs to the family of integral plasma membrane proteins; solute carrier family 12 ,the electrically silent, cation-coupled chloride co-transporter family that mediates electroneutral symport of Na+, K+, and Cl with a stochiometry of 1:1:2, respectively. NKCC2 structure contains a central hydrophobic domain made up of 12 putative transmembrane spanning regions and a long hydrophilic loop connects transmembrane segments 7 and 8 and contains two glycosylation sites. The hydrophobic domain is flanked by a short amino- and a long carboxyl-terminal domain that are located within the cell.

NKCC2 encoded by the Slc12a1 gene shows at least six alternatively spliced variants and has the ability to form dimmers. NKCC2 activity and expression can be found in the medullary and cortical thick ascending limb of Henle's loop is the nephron and in macula densa cells.

NKCC2 allows Na+ and Cl entry from the tubule lumen into the thick ascending limb of  Henle's loop, a process that allows reabsorbing 25–30% of the NaCl filtered by the glomeruli. In addition, NKCC2 can sense luminal Cl concentration and initiates the signal for tubuloglomerular feedback in the specialized cells of the macula densa. Thus, NKCC2 activity is essential for salt conservation and for acute and chronic regulation of water balance.

Mutations in the human NKCC2 gene causes Bartter syndrome type 1 characterized by severe natriuresis hypercalciuria and magnesuria, the inability to concentrate urine, leading to hyponatremia, hypochloremia, metabolic alkalosis, and low blood pressure. Opposite to Bartter's syndrome, enhanced activity of NKCC2 NaCl is connected with human and animal hypertension.

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: 08/06/2021

Alomone Labs is pleased to offer a highly specific antibody directed against an epitope of rat NKCC2 transporter. Anti-NKCC2 (SLC12A1) (extracellular) Antibody (#ANT-072). The antibody recognizes an extracellular epitope and is thus ideal for detecting NKCC2 in living cells. It has been designed to recognize NKCC2 from rat, human and mouse samples.

For research purposes only, not for human use

Applications

Specifications

Scientific Background

Citations

Citations
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