Bone Remodeling and Osteoclasts:
Bones are complex, living organs that distinguish vertebrates from
other animals. Bones serve multiple purposes in the human body,
including assisting in mobility, serving as warehouses for the
production of blood cells, and functioning as reservoirs for storage
and release of essential minerals like calcium. While the bone might appear rigid and inert, it is in fact
a dynamic organ that undergoes constant remodeling (Crockett et al., 2011). Estimates show that
approximately 10% of the total human bone mass is remodeled every year (Jones et al., 2002). Bone
remodeling, also termed bone metabolism, is a life-long process required for many skeletal functions
including bone growth, fracture healing, tooth eruption, and maintenance of steady state calcium levels
in the blood... more
Kingfisher Biotech has a growing portfolio of RANKL proteins. We are committed to accelerating basic veterinary and animal model research by developing and commercializing reagents to various species, many of which have been previously under-served. Our products span over a dozen species (bovine, canine, caprine, catfish, chicken, dolphin, equine, feline, guinea pig, hamster, human, mouse, ovine, rabbit, rat, swine, and turkey). Our product offering includes recombinant proteins, polyclonal antibodies, monoclonal antibodies, Do-It-Yourself ELISAs, and ELISA kits.
Showing posts with label RANKL. Show all posts
Showing posts with label RANKL. Show all posts
Thursday, May 14, 2015
Friday, February 22, 2013
RANKL Bioactivity
An independent laboratory with expertise in osteoclast cell culture has confirmed the bioactivity of Kingfisher Biotech's mouse RANKL/TNFSF11 protein (RP0437M-005, RP0437M-025, and RP0437M-100).
Receptor Activator of Nuclear factor Kappa-B Ligand (RANKL), also known as Tumor necrosis factor ligand superfamily member 11 (TNFSF11), TNF-related activation-induced cytokine (TRANCE), osteoprotegerin ligand (OPGL), and osteoclast differentiation factor (ODF) is a member of the tumor necrosis factor (TNF) ligand family. RANKL regulates bone remodeling and the development of the thymus, lymph nodes and mammary glands. Overproduction of RANKL is implicated in a variety of degenerative bone diseases, such as rheumatoid arthritis and psoriatic arthritis.
Kingfisher Biotech’s Mouse RANKL/TNFSF11 activity was accessed and shown to be active and comparable with other commercially available Human RANKL protein. This work was done by an independent lab with expertise in osteoclast cell culture. Both samples of RANKL were reconstituted with 0.1% BSA in PBS prior to assay. Kingfisher Biotech’s RANKL protein concentration was determined by BCA. Protein determination of the competitor’s protein is unknown. Kingfisher Biotech’s mouse RANKL was expressed in yeast so it is naturally endotoxin-free and post-translationally modified. Human RANKL purchased from Competitor A was produced in E. coli.
Method
RAW 264.7 cells (1.3 × 104/ml) were cultured in 24-well plates for 5 days in the presence of RANKL (100 ng/ml) and M-CSF (10 ng/ml). The culture media was changed every alternate day and replaced with fresh media and cytokines. At the end of the culture period, cells were fixed in 2% glutaraldehyde in PBS for 20 min and stained for TRAP activity (Sigma, 387A). TRAP positive multinucleated cells containing three or more nuclei were scored microscopically. The experiment was repeated 15 days after the initial experiment and yielded similar results.
Assay Results
Receptor Activator of Nuclear factor Kappa-B Ligand (RANKL), also known as Tumor necrosis factor ligand superfamily member 11 (TNFSF11), TNF-related activation-induced cytokine (TRANCE), osteoprotegerin ligand (OPGL), and osteoclast differentiation factor (ODF) is a member of the tumor necrosis factor (TNF) ligand family. RANKL regulates bone remodeling and the development of the thymus, lymph nodes and mammary glands. Overproduction of RANKL is implicated in a variety of degenerative bone diseases, such as rheumatoid arthritis and psoriatic arthritis.
Kingfisher Biotech’s Mouse RANKL/TNFSF11 activity was accessed and shown to be active and comparable with other commercially available Human RANKL protein. This work was done by an independent lab with expertise in osteoclast cell culture. Both samples of RANKL were reconstituted with 0.1% BSA in PBS prior to assay. Kingfisher Biotech’s RANKL protein concentration was determined by BCA. Protein determination of the competitor’s protein is unknown. Kingfisher Biotech’s mouse RANKL was expressed in yeast so it is naturally endotoxin-free and post-translationally modified. Human RANKL purchased from Competitor A was produced in E. coli.
Method
RAW 264.7 cells (1.3 × 104/ml) were cultured in 24-well plates for 5 days in the presence of RANKL (100 ng/ml) and M-CSF (10 ng/ml). The culture media was changed every alternate day and replaced with fresh media and cytokines. At the end of the culture period, cells were fixed in 2% glutaraldehyde in PBS for 20 min and stained for TRAP activity (Sigma, 387A). TRAP positive multinucleated cells containing three or more nuclei were scored microscopically. The experiment was repeated 15 days after the initial experiment and yielded similar results.
Assay Results
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