Klotho beta Antibody [Unconjugated]

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.

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.

Effects of FGF21 on beta ‐klotho expression and oxidative stress in EPCs. After FGF21 treatment, cells were (A) stained with an beta ‐klotho antibody, and (B) protein expression was analysed by Western blotting. (C) The effects of FGF21 on EPC viability were analysed by the CCK‐8 assay. (D) EPCs were treated with the indicated concentration of FGF21 for 12 h and 600 µM H2O2 for 1 h. Intracellular ROS levels were measured using a Fluorometric Intracellular ROS Kit. The results are expressed as the mean ± SEM of five separate experiments run in triplicate (*p < 0.01 vs. control, #p < 0.01 vs. H2O2 only) Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35307922), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Klotho beta Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human Klotho beta
  Phe53-Leu997
  Accession # Q86Z14
• 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: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: KLB
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Western Blot 0.5 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.2 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Klotho beta Antibody [Unconjugated]

• betaKlotho
• beta-klotho
• BKL
• KLB
• klotho beta like
• Klotho beta
• klotho beta-like protein
• MGC142213

Background

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.

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Klotho beta Antibody (AF5889)(4)

Publications using AF5889

• WP Huang, CY Chen, TW Lin, CS Kuo, HL Huang, PH Huang, SJ Lin Fibroblast growth factor 21 reverses high-fat diet-induced impairment of vascular function via the anti-oxidative pathway in ApoE knockout mice Journal of Cellular and Molecular Medicine, 2022-03-20;0(0):. 2022-03-20 [PMID: 35307922] (ICC, Human)
• F Li, X Li, Z Li, W Ji, S Lu, W Xia betaKlotho is identified as a target for theranostics in non-small cell lung cancer Theranostics, 2019-10-12;9(25):7474-7489. 2019-10-12 [PMID: 31695781] (Western Blot, Human)
• Gao R, Kanasaki K, Li J et al. Beta klotho is essential for the anti-endothelial mesenchymal transition effects of N-acetyl-seryl-aspartyl-lysyl-proline FEBS Open Bio. NULL [PMID: 30972974]
• Ming AY, Yoo E, Vorontsov EN, Altamentova SM, Kilkenny DM, Rocheleau JV 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. J. Biol. Chem., 2012-04-20;287(24):19997-20006. 2012-04-20 [PMID: 22523080] (Western Blot, Human)

Bioinformatics

• Gene Symbol: KLB
• Uniprot:
  • Q86Z14()