Human Klotho  beta Antibody

Recombinant Monoclonal Antibody.
  
  • Species Reactivity
    Human
  • Specificity
    Detects human Klotho beta in direct ELISAs.
  • Source
    Recombinant Monoclonal Rabbit IgG Clone # 1025C
  • Purification
    Protein A or G purified from cell culture supernatant
  • Immunogen
    Mouse myeloma cell line, NS0-derived recombinant human Klotho beta with a C-terminal 10 His tag.
    Phe53-Leu997
    Accession # Q86Z14
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Flow Cytometry
    0.25 µg/106 cells
    See below
  • Immunocytochemistry
    3-25 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Klotho  beta in HepG2 Human Cell Line by Flow Cytometry. HepG2 human hepatocellular carcinoma cell line was stained with Rabbit Anti-Human Klotho  beta Monoclonal Antibody (Catalog # MAB58891, filled histogram) or isotype control antibody (Catalog # AB-105-C, open histogram), followed by Allophycocyanin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0111). View our protocol for Staining Membrane-associated Proteins.
Immunocytochemistry
Klotho  beta in HepG2 Human Cell Line. Klotho  beta was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Rabbit Anti-Human Klotho  beta Monoclonal Antibody (Catalog # MAB58891) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Klotho beta

Klotho beta, a divergent structural member of the glycosidase I superfamily, is expressed primarily in the liver and pancreas, with lower expression in adipose tissue (1, 2). Like Klotho, Klotho beta facilitates binding between FGF19 subfamily members and their receptors via formation of a ternary complex (3). The Klotho beta mediated interaction of human FGF19 (mouse FGF15) with FGF Receptor 4 in the liver negatively regulates bile acid synthesis by controlling the secretion of two key bile acid synthase genes, cholesterol 7-alpha hydroxylase (Cyp7a1) and sterol 12-alpha hydroxylase (Cyp8b1) (2-5). Klotho beta is also a cofactor for the interaction of FGF21 with FGF Receptor 1c in adipocytes, which allows FGF21 to stimulate GLUT1 expression, upregulating adipocyte insulin-dependent glucose uptake (2-4, 6). The 1043 amino acid (aa) type I transmembrane protein is composed of a 51 aa signal sequence, a 943 aa extracellular domain (ECD) containing two glycosidase-like regions, a 21 aa transmembrane domain, and 28 aa intracellular tail. Since Klotho-related proteins lack critical active site Glu residues present in beta -glycosidases, it was initially unclear whether they were functional enzymes (1, 7). However, glucuronidase activity has since been demonstrated for Klotho, indicating that physiologically relevant enzymatic activity for Klotho beta is also possible (8). The extracellular domain shares 79%, 87%, 87% and 67% identity with mouse, equine, canine and rat Klotho beta, respectively. The low identity with rat reflects aa discordance within rodent ECD.

  • References:
    1. Mian, I. S. (1998) Blood Cells Mol. Dis. 24:83.
    2. Kurosu, H. and M. Kuro-o (2009) Mol. Cell. Endocrinol. 299:72.
    3. Ito, S. et al. (2005) J. Clin. Invest. 115:2202.
    4. Kurosu, H. et al. (2007) J. Biol. Chem. 282:26687.
    5. Lin, B. C. et al. (2007) J. Biol. Chem. 282:27277.
    6. Ogawa, Y. et al. (2007) Proc. Natl. Acad. Sci USA 104:7432.
    7. Chang, Q. et al. (2005) Science 310:490.
    8. Goetz, R. et al. (2007) Mol. Cell. Biol. 27:3417.
  • Entrez Gene IDs:
    152831 (Human); 83379 (Mouse); 289625 (Rat)
  • Alternate Names:
    betaKlotho; beta-klotho; BKL; KLB; klotho beta like; Klotho beta; klotho beta-like protein; MGC142213
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Donkey Anti-Rabbit IgG (H+L) Antibody

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