Recombinant Human Klotho His-tag Protein, CF

When Recombinant Human FGF-23 (Catalog # 2604-FG) is immobilized at 5 μg/mL, 100 μL/well, Recombinant Human Klotho Mucin Stalk Chimera His-tag (Catalog # 10308-KL) binds with an ED50 of 0.2‑1.8 μg/mL.

2 μg/lane of Recombinant Human Klotho Mucin Stalk Chimera His-tag (Catalog # 10308-KL) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 110-140 kDa.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human Klotho His-tag Protein, CF Summary

• Additional Information: Mucin Stalk Chimera
• Details of Functionality: Measured by its binding ability in a functional ELISA. When Recombinant Human FGF-23 (Catalog # 2604-FG) is immobilized at 5 µg/mL (100 µL/well), Recombinant Human Klotho Mucin Stalk Chimera His-tag (Catalog # 10308-KL) binds with an ED₅₀ of 0.2-1.8 μg/mL.
• Source: Chinese Hamster Ovary cell line, CHO-derived human Klotho protein
  

  Human Klotho (Glu34-His549) Accession # Q9UEF7	Human Fractalkine Mucin-like Stalk (Phe103-Thr338) Accession # P78423	6-His tag
  N-terminus		C-terminus

• Accession #: Q9UEF7 (Klotho) & P78423 (Fractalkine Mucin-like S
• N-terminal Sequence: Glu34
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 84 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 110-140 kDa, under reducing conditions

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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 500 μg/mL in PBS.

Notes

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

Alternate Names for Recombinant Human Klotho His-tag Protein, CF

• EC 3.2.1
• EC 3.2.1.31
• KL
• Klotho

Background

Klotho, also called alpha-Klotho ( alpha -Klotho), is the founding member that along with beta -Klotho and gamma -Klotho, form the Klotho family within the glycosidase-1 superfamily (1, 2). alpha -Klotho is a type I transmembrane protein consisting of a large extracellular domain (ECD) containing glycosidase-like domains (KL1 and KL2), a single transmembrane domain and a short intracellular domain. Alternative mRNA splicing of the ECD of alpha -Klotho results in a circulating protein known as soluble alpha -Klotho (s‑Klotho), which has been detected in both humans and mice (3, 4). In addition to the s-Klotho form, a 130 kDa form found in plasma and cerebrospinal fluid and a prominent intracellular 120 kDa form of alpha -Klotho have also been identified (3, 4). The mature ECD of full length human alpha -Klotho shares 87% and 90% identity with mouse and rat alpha -Klotho, respectively. Due to highly conserved sequences between alpha -Klotho forms, it is difficult to differentiate s-klotho from the other short forms in vivo (5). Although alpha -Klotho was identified ~20 years ago, its function remains incompletely understood. alpha -Klotho shows weak glucuronidase activity which activates the renal ion channel TRPV5 to reabsorb urinary calcium (10). alpha -Klotho acts as a cofactor for interaction of FGF-23 with FGF R1 (6). This interaction negatively regulates 1 alpha -hydroxylase, the rate-limiting enzyme in the synthesis of 1,25(OH)2D3 (vitamin D) (7). s-Klotho functions as a hormonal factor and is involved in anti-aging, anti-oxidation, modulation of ion-transport, and Wnt signaling (8). Both alpha -Klotho and beta -Klotho are cofactors for FGF19 binding (9). The phenotype of alpha ‑Klotho-deficient mice resembles premature aging, including arteriosclerosis, osteoporosis, skin atrophy, infertility, emphysema and premature death (2). alpha -Klotho deficient mice show severe hyperphosphatemia and ectopic calcification of soft tissues due to excess vitamin D (2-7). Conversely, excess alpha -Klotho extends lifespan (6).

1. Nabeshima, Y. (2006) Sci. Aging Knowl. Environ. 8:pe11.
2. Kuro-o, M. et al. (1997) Nature 390:45.
3. Shiraki-Iida, T. et al. (1998) FEBS Lett. 424:6.
4. Imura, A. et al. (2004) FEBS Lett. 565:143.
5. Yuechi, X. et al. (2015) Endocr Rev. 36:174.
6. Kurosu, H. et al. (2006) J. Biol. Chem. 281:6120.
7. Tsujikawa, H. et al. (2003) Mol. Endocrinol. 17:2393.
8. Hu, M. et al. (2012) Adv Exp Med Biol. 728:126.
9. Wu, X. et al. (2007) J. Biol. Chem. 282:29069.
10. Chang, Q. et al. (2005) Science 310:490.

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Bioinformatics

• Uniprot:
  • Q9UEF7 (Klotho) & P78423 (Fractalkine Mucin-like S()