FREE soluble RANKL ELISA
- Product Details
- Instructions For Use
- Validation Data
- Additional Information
- References, Applications
|Method:||Sandwich ELISA, HRP/TMB, 12x8-well strips|
|Sample type:||Serum, heparin plasma|
|Standard range:||0-2 pmol/l (7 serum based standards)|
|Standard points:||0/0.0625/0.125/0.25/0.5/1/2 pmol/l|
|Controls:||2 serum based controls|
|Sample size:||150 µl / well|
|Incubation time:||2 h / overnight / 1 h / 30 min|
|Unit conversion:||1 pg/ml = 0.05 pmol/l (MW: 20 kDa, monomer)|
LOD: 0.01 pmol/l (0 pmol/l + 3 SD); LLOQ: 0.008 pmol/l
Intra-assay (n=5) ≤ 5%, Inter-assay (n=12) ≤ 3%
The mean recovery of endogenous free sRANKL (spike: 0.15 pmol/l; 0.34 pmol/l) is:
Serum: 89%; 85%
Heparin plasma: 100%; 98%
Dilution linearity of endogenous free sRANKL with a low measuring serum as dilution medium is:
Serum: 1+1 dilution = 112%, 1+3 dilution = 121%
Heparin plasma: 1+1 dilution = 121%
The sequence homology to various primates is >95%. It is likely that the assay can be used for these species. Internal validations have not been carried out.
Values from apparently healthy individuals:
Median (serum, n=32): 0.14 pmol/l.
It is recommended to establish the normal range for each laboratory.
Principle of the assay:
Manual ELISAs easily adaptable for automation!
INSTRUCTIONS FOR USE
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BI-20462 - free sRANKL ELISA Validation Data (S/R, dilution linearity, precision, ...)
BI-20462 - free sRANKL ELISA Validation Data by external CRO
RANKL, the receptor activator of nuclear factor kappa B ligand, a member of the tumor necrosis factor (TNF) family (http://www.uniprot.org/uniprot/O14788), is the main stimulatory factor for the formation of mature osteoclasts and is essential for their survival. RANKL activates its specific receptor RANK, located on osteoclasts and dendritic cells.
The effects are counteracted by OPG which acts as an endogenous soluble receptor antagonist (see: BI-20403 – OPG ELISA).
The major source of RANKL are osteocytes, former osteoblasts that become embedded within the mineralized bone matrix. RANKL is a ~35 kDa type II transmembrane-type protein and is cleaved to release a soluble biologically active product that forms a homotrimer.
RANKL and its specific receptor RANK are not only key regulators of bone remodeling but also play an essential role in immunobiology, e.g. lymph node formation, establishment of the thymic microenviroment, mammary gland development during pregnancy, bone metastasis in cancer and sex-hormone, progestin-driven breast cancer, thermoregulation, and finally in the development of type 2 diabetes mellitus.
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