Anti-NBCe1 (extracellular) Antibody

Electrogenic Sodium Bicarbonate Cotransporter 1, NBC1, SLC4A4
    Cat #: ANT-075
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: m, r
    Immunogen
    Peptide (C)KE(S)VKFGGKLVGNN, corresponding to amino acid residues 672-685 of rat NBCe1 (Accession Q9JI66). 2nd extracellular loop, based on the 10 transmembrane topology.
    Accession (Uniprot) Number Q9JI66
    Gene ID 84484
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Rat - 13/14 amino acid residues identical; mouse - 12/14 amino acid residues identical.
    Purity Affinity purified on immobilized antigen.
    Formulation Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
    Specificity Not recommended for human samples. The antibody will recognize all known rat splice variants
    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).
    Control antigen storage before reconstitution Lyophilized powder can be stored intact at room temperature for 2 weeks. For longer periods, it should be stored at -20°C.
    Control antigen reconstitution 100 µl double distilled water (DDW).
    Control antigen storage after reconstitution -20°C.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ic, ih, lci, wb
    May also work in: ifc, ip
    Western blot
    Western blot analysis of rat brain lysate:
    1. Anti-NBCe1 (extracellular) Antibody (#ANT-075), (1:200).
    2. Anti-NBCe1 (extracellular) Antibody, preincubated with the control peptide antigen.
    Western blot analysis of mouse liver membranes:
    1. Anti-NBCe1 (extracellular) Antibody (#ANT-075), (1:400).
    2. Anti-NBCe1 (extracellular) Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Expression of NBCe1 in rat lateral septum
    Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using Anti-NBCe1 (extracellular) Antibody (#ANT-075), (1:400). NBCe1 staining (red) is detected in blood vessel outlines (vertical arrow) in the lateral septum (LS) and along the wall of the lateral ventricle (LV) (horizontal arrow).  DAPI is used as the counterstain (blue).
    Live cell imaging / Immunocytochemistry
    Expression of NBCe1 in rat PC12 cells
    Immunocytochemical staining of live intact rat PC12 pheochromocytoma cells. A. Extracellular staining of cells with Anti-NBCe1 (extracellular) Antibody (#ANT-075), (1:100), followed by goat anti-rabbit-AlexaFluor-594 secondary antibody (red). B. Live image of the cells. C. Merge of the two images.
    References
    1. Romero, M.F. et al. (2013) Mol. Aspects Med. 34,159.
    2. Abuladze, N. et al.(2000) Gene 251, 109.
    3. Zhu, Q. et al. (2010) J. Biol. Chem. 285, 13416.
    4. Kao, L. et al. (2008) J. Biol. Chem. 283, 26782.
    5. Igarashi, T. et al. (1999) Nat. Genet. 23, 264.
    Scientific background

    SLC4 membrane transporter proteins play important roles in kidney acid–base regulation through their transport of bicarbonate (or carbonate), Na+, Cl-, and (possibly NH4+)1. These transporters differ in their substrate (Na+, Cl-) dependence, charge transport stoichiometry, cell-type and developmental expression, functional regulation, and protein–protein interactions. In mammals, SLC4 proteins are encoded by 10 different genes that share protein sequence homology and are grouped according to their functional properties.

    In humans, the electrogenic sodium bicarbonate cotransporter NBCe-1 is encoded by the SLC4A4 gene2. NBCe1-A has a large N-terminal cytoplasmic region, a lipid embedded transmembrane region, and a C-terminal cytoplasmic tail3. The N-terminal cytoplasmic region is tightly folded and is predicted to form a domain structure, unlike the freely aqueous accessible C-terminal cytoplasmic tail. NBCe1-A is composed of either 10 or 14 transmembrane domains. Based on the 10 transmembrane topology, a large extracellular loop (EL) is present between TM3 and TM4 and contains two glycosylated sites. The oligomeric state of the cotransporter is dimeric and each monomeric subunit has independent transport activity4.

    Mutations in NBCe1 cause autosomal recessive proximal renal tubular acidosis (pRTA). Patients with NBCe1 mutations have severe pRTA, growth and mental retardation, basal ganglia calcification, cataracts, corneal opacities (band keratopathy), glaucoma, elevated serum amylase and lipase, and defects in the enamel consistent with amelogenesis imperfect3. Mice with loss of NBCe1 have a more severe phenotype with marked volume deletion and decreased survival5.

    Application key:

    CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, 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: 30/10/2018

    Alomone Labs is pleased to offer a highly specific antibody directed against an extracellular epitope of rat NBCe1. Anti-NBCe1 (extracellular) Antibody (#ANT-075) can be used in western blot analysis, immunohistochemistry and live cell imaging applications. The antibody was used to probe NBCe1 in living rat PC12 cells. It has been designed to recognize NBCe1 from rat and mouse samples and is not recommended to use in human samples.

    For research purposes only, not for human use
    Citations
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