Recombinant Human Klotho His-tag Protein, CF

Catalog #: 10308-KL Datasheet
Mucin Stalk Chimera
Catalog # Availability Size / Price Qty
10308-KL-050
Recombinant Human Klotho His-tag Protein Binding Activity
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Recombinant Human Klotho His-tag Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
Activity
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-terminusC-terminus
N-terminal Sequence
Analysis
Glu34
Predicted Molecular Mass
84 kDa
SDS-PAGE
110-140 kDa, under reducing conditions

Product Datasheets

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10308-KL

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

10308-KL

Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 500 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it 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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Scientific Data

Binding Activity Recombinant Human Klotho His-tag Protein Binding Activity View Larger

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.

SDS-PAGE Recombinant Human Klotho His-tag Protein SDS-PAGE View Larger

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.

Background: Klotho

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).

References
  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.
Entrez Gene IDs
9365 (Human); 16591 (Mouse)
Alternate Names
EC 3.2.1; EC 3.2.1.31; KL; Klotho

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