Human Klotho  beta Antibody

(1 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human Klotho beta in direct ELISAs and Western blots. In direct ELISAs, approximately 35% cross-reactivity with recombinant mouse Klotho beta is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Klotho beta
    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
  • Western Blot
    0.5 µg/mL
    See below
  • Immunocytochemistry
    5-15 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 Human Klotho  beta by Western Blot. Western blot shows lysates of Hep3B human hepatocellular carcinoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human Klotho  beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5889) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Klotho  beta at approximately 130 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
Klotho  beta in HepG2 Human Cell Line. Klotho  beta was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Goat Anti-Human Klotho  beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5889) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) 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
    Sterile PBS to a final concentration of 0.2 mg/mL.
  • 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
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citations: Showing 1 - 1

  1. Dynamics and Distribution of Klotho beta (KLB) and fibroblast growth factor receptor-1 (FGFR1) in living cells reveal the fibroblast growth factor-21 (FGF21)-induced receptor complex.
    Authors: Ming AY, Yoo E, Vorontsov EN, Altamentova SM, Kilkenny DM, Rocheleau JV
    J. Biol. Chem., 2012;287(24):19997-20006.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
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