Detection of Human Klotho beta by Western Blot.
Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line and Hep3B human hepatocellular carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Klotho beta Monoclonal Antibody (Catalog # MAB5889) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). Specific bands were detected for Klotho beta at approximately 130-140 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Klotho beta by Simple WesternTM.
Simple Western lane view shows lysates of HepG2 human hepatocellular carcinoma cell line and Hep3B human hepatocellular carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Klotho beta at approximately 177/170 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human Klotho beta Monoclonal Antibody (Catalog # MAB5889). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
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 (b-Klotho or KLB) is a 125-135 kDa member of the KL family, Glycosyl Hydrolase 1 superfamily of proteins. It plays key role in the biology of the endocrine FGFs (FGF-15/19; 21; 23). KLB appears to serve as a signaling cofactor for select FGFRs, including FGFR-1c, -2c, -3c and FGFR4. The cofactor is known to constitutively interact with FGFRs, promote FGF binding to their appropriate FGFRs, and to bind directly to FGF-19 and FGF-21. Notably, FGF-23 appears to utilize a-Klotho/KL for its FGFR signaling cofactor, and FGF-1, while not dependent upon KLB for signaling, can utilize KBL to potentiate certain activities. KLB is expressed on select cells, including adipocytes and hepatocytes. Human KLB is a 1044 amino acids (aa) type III transmembrane (TM) protein (i.e.-a type I TM protein lacking a signal sequence). It contains a 996 aa extracellular region (aa 1-996) and a 27 aa cytoplasmic domain (aa 1018-1044). The extracellular region contains two glycosyl hydrolase domains (aa 77-508 and 517-967) plus eleven potential N-linked glycosylation sites. Neither hydrolase domain is active as they lack a key Glu residue needed for activity. Over aa 53-997, human KLB shares 80% aa sequence identity with mouse KLB. And relative to KL/a-Klotho, human KLB and KL share just 44% aa sequence identity over the entire length of the molecules.
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