Human Klotho  beta Antibody

Recombinant Monoclonal Antibody
Catalog # Availability Size / Price Qty
MAB58891
MAB58891-SP
Detection of Klotho  beta  in HepG2 Human Cell Line by Flow Cytometry.
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Human Klotho  beta Antibody Summary

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 either lyophilized or 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.

Scientific Data

Flow Cytometry Detection of Klotho  beta  antibody in HepG2 Human Cell Line antibody by Flow Cytometry. View Larger

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  antibody in HepG2 Human Cell Line by Immunocytochemistry (ICC). View Larger

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.

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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); 102147155 (Cynomolgus Monkey)
Alternate Names
betaKlotho; beta-klotho; BKL; KLB; klotho beta like; Klotho beta; klotho beta-like protein; MGC142213

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