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MABS1237 Anti-NKCC1/2 Antibody, clone T4

This mouse monoclonal Anti-NKCC1/2 Antibody, clone T4, Cat. No. MABS1237 detects levels of NKCC1 & NKCC2, and has been published and validated for use in Electron Microscopy, Immunocytochemistry, Immunofluorescence, Immunoprecipitation, and Western Blotting.

Anti-NKCC1/2 Antibody, clone T4 MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Research Category: Signaling Gene Symbol: SLC12A1, SLC12A2, NKCC2, NKCC1 UniProt Number: P55011
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MABS1237
100 μg  
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      Overview

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      Key Specifications Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      B, R, Av, H, Gr, M, Rb, F, Ft, Shk, CaEM, ICC, IF, IP, WBMPurifiedMonoclonal Antibody
      Description
      Catalogue NumberMABS1237
      DescriptionAnti-NKCC1/2 Antibody, clone T4
      Alternate Names
      • Solute carrier family 12 member 1
      • Solute carrier family 12 member 2
      • Bumetanide-sensitive sodium-(potassium)-chloride cotransporter 2
      • Bumetanide-sensitive sodium-(potassium)-chloride cotransporter 1
      • Basolateral Na-K-Cl symporter
      Background InformationSolute carrier family 12 member 1 (UniProt Q13621; a.k.a. Bumetanide-sensitive sodium-(potassium)-chloride cotransporter 2) and solute carrier family 12 member 2 (UniProt P55011; a.k.a. Basolateral Na-K-Cl symporter, Bumetanide-sensitive sodium-(potassium)-chloride cotransporter 1) are encoded by the SLC12A1 (a.k.a. NKCC2; Gene ID 6557) and SLC12A2 (a.k.a. NKCC1; Gene ID 6558) gene, respectively, in human. NKCC1 (SLC12A2) and NKCC2 (SLC12A1) belong to the subfamily of cation-chloride cotransporters (CCCs) that mediate electroneutral transport of sodium, potassium and chloride across the plasma membrane and are inhibited by bumetanide and furosemide-type of compounds. NKCCs play an important role in cell volume regulation, where NKCC2 is involved in reabsorption of salt and osmotically-obliged water in renal epithelial cells, while NKCC1 has a major impact on intracellular chloride ion concentration ([Cl-]i) in vascular smooth muscle cells (VSMCs) and neuronal cells. In primary hypertension, NKCC1 activity is increased in VSMCs and paraventricular nucleus (PVN) neurons, leading to elevated peripheral resistance in systemic circulation and activation of sympathetic nerve system (SNS), respectively. Human NKCC1 and NKCC2 are 12-transmembrane proteins having their N- and C-terminal ends exposed at the cytoplasmic side (a.a. 1-285 and 921-1212 of NKCC1; a.a. 1-177 and 814-1099 of NKCC2).
      References
      Product Information
      FormatPurified
      PresentationPurified mouse monoclonal IgG1κ in buffer containing 0.1 M Tris-Glycine (pH 7.4) 150 mM NaCl with 0.05% sodium azide
      Quality LevelMQ100
      Applications
      ApplicationThis mouse monoclonal Anti-NKCC1/2 Antibody, clone T4, Cat. No. MABS1237 detects levels of NKCC1 & NKCC2, and has been published and validated for use in Electron Microscopy, Immunocytochemistry, Immunofluorescence, Immunoprecipitation, and Western Blotting.
      Key Applications
      • Electron Microscopy
      • Immunocytochemistry
      • Immunofluorescence
      • Immunoprecipitation
      • Western Blotting
      Application NotesImmunohistochemistry Analysis: A 1:3,000 dilution from a representative lot detected NKCC in human prostate cancer, as well as normal colon, small intestine, and testis tissue sections.

      Electron Microscopy Analysis: A representative lot detected NKCC immunoreactivity in 0.5% glutaraldehyde and 4% paraformaldehyde gerbil inner ear tissue sections (Crouch, J.J., et al. (1997). J. Histochem. Cytochem. 45(6):773-778).

      Immunofluorescence Analysis: A representative lot detected NKCC-positive cells by fluorescent immunohistochemistry staining of 4% paraformaldehyde- or in 2.5% glutaraldehyde-fixed, paraffin-embedded fish gill filaments (Brauer, P.R., et al. (2005). Anat. Rec. A Discov. Mol. Cell. Evol. Biol. 285(1):600-609).

      Immunofluorescence Analysis: Representative lots detected brain tissue NKCC immunoreactivity by fluorescent immunohistochemistry staining of free-floating mouse and rat brain sections (Liu, F., et al. (2010). Exp. Neurol. 224(2):356-361; Yan, Y., et al. (2001). J. Cereb. Blood Flow Metab. 21(6):711 721).

      Immunofluorescence Analysis: A representative lot detected NKCC immunolocalization by fluorescent immunohistochemistry staining of MeOH (80%)/DMSO (20%) fixed rat kidney cryosections (Ginns, S.M., et al. (1996). J. Am. Soc. Nephrol. 7(12):2533-2542).

      Immunofluorescence Analysis: A representative lot immunostained basolateral margin of the acinar cells, but not the apical pole or cell interior by fluorescent immunohistochemistry staining of PLP-fixed (2% paraformaldehyde, 75 mM lysine, 10 mM sodium periodate) rabbit parotid gland cryosections (Lytle, C., et al. (1995). Am. J. Physiol. 269(6 Pt 1):C1469-C1505).

      Immunohistochemistry Analysis: A representative lot detected NKCC immunoreactivity by immunohistochemistry staining of 4% paraformaldehyde-fixed free-floating rat brain sections (Yan, Y., et al. (2001). J. Cereb. Blood Flow Metab. 21(6):711 721).

      Immunohistochemistry Analysis: A representative lot detected NKCC immunoreactivity in 10% formalin-fixed, Paraplast Plus-embedded gerbil kidney and inner ear tissue sections (Crouch, J.J., et al. (1997). J. Histochem. Cytochem. 45(6):773-778).

      Immunohistochemistry Analysis: A representative lot detected Na-K-Cl cotransporters in serous acinar cells, but not along striated ducts in PLP-fixed (2% paraformaldehyde, 75 mM lysine, 10 mM sodium periodate) rabbit parotid gland cryosections (Lytle, C., et al. (1995). Am. J. Physiol. 269(6 Pt 1):C1469-C1505).

      Immunoprecipitation Analysis: Representative lot immunoprecipitated denatured, but not non-denatured human and canine Na-K-Cl cotransport proteins (Lytle, C., et al. (1995). Am. J. Physiol. 269(6 Pt 1):C1469-C1505; Haas, M., et al. (1995). J. Biol. Chem. 270(48):28955 28961).

      Western Blotting Analysis: A representative lot detected exogenously expressed ferret NKCC1 & NKCC2A in lysates from transfected HEK293 cells (Hannemann, A., and Flatman, P.W. (2011). PLoS One. 6(3):e17992).

      Western Blotting Analysis: Representative lots detected mouse, rat, fish, and bovine NKCC proteins (Liu, F., et al. (2010). Exp. Neurol. 224(2):356-361; Brauer, P.R., et al. (2005). Anat. Rec. A Discov. Mol. Cell. Evol. Biol. 285(1):600-609; Abbruscato, T.J., et al. (2004). J. Pharmacol. Exp. Ther. 310(2):459-468; Yan, Y., et al. (2001). J. Cereb. Blood Flow Metab. 21(6):711 721).

      Western Blotting Analysis: A representative lot detected Na-K-Cl cotransport protein (~160-195 kDa) in membrane extracts from shark rectal gland, mouse medullary thick ascending limb, canine kidney, duck salt gland, human colonic T84 cells, and rabbit parotid salivary gland. N-glycosylase F treatment of the extracts reduced the sizes of target bands (Lytle, C., et al. (1995). Am. J. Physiol. 269(6 Pt 1):C1469-C1505).

      Note: Clone T4 epitope is not exposed on the surface of Na-K-Cl cotransport protein in its native conformation. Denature NKCC with the addtion of SDS to a final concentration of 1% in test samples, then diluting the denatured samples into 6 v
      Biological Information
      ImmunogenRecombinant human NKCC1 C-terminal fragment.
      CloneT4
      ConcentrationPlease refer to lot specific datasheet.
      HostMouse
      SpecificityClone T4 recognizes both NKCC1 and NKCC2 by targeting a C-terminal region epitope exposed only under denaturing condition, and unaccessible for antibody binding when NKCC assumes its native conformation (Lytle, C., et al. (1995). Am. J. Physiol. 269(6 Pt 1):C1469-C1505).
      IsotypeIgG1κ
      Species Reactivity
      • Bovine
      • Rat
      • Avian
      • Human
      • Gerbil
      • Mouse
      • Rabbit
      • Fish
      • Ferret
      • Shark
      • Canine
      Species Reactivity NoteAvian, Bovine, Canine, Ferret, Fish, Gerbil, Human, Mouse, Rabbit, Rat, Shark.
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Gene Symbol
      • SLC12A1
      • SLC12A2
      • NKCC2
      • NKCC1
      Purification MethodProtein G purified.
      UniProt Number
      Molecular Weight~165 kDa observed. Target bands appear larger than the calculated molecular weights of 131.4/129.7 kDa (human NKCC1 isoform 1/2), 121.5/121.4 kDa (human NKCC2 isoform A/F), 131.0 kDa (mouse NKCC1), 120.4/120.3/120.3 kDa (mouse NKCC2 isoform A/B/F), and 130.4/120.6 kDa (rat NKCC1/NKCC2) due to glycosylation. Uncharacterized bands may be observed in some lysate(s).
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceEvaluated by Western Blotting in HeLa cell lysate.

      Western Blotting Analysis: 0.5 µg/mL of this antibody detected NKCC in 10 µg of HeLa cell lysate.
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsStable for 1 year at 2-8°C from date of receipt.
      Packaging Information
      Material Size100 μg
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalog Number GTIN
      MABS1237 04054839035340