CLIA Kit for Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL)

CD254; TNFSF11; ODF; OPGL; TRANCE; HRANKL2; SOdf; Tumor Necrosis Factor(ligand)superfamily Member 11; TNF-related activation-induced cytokine

Specificity

This assay has high sensitivity and excellent specificity for detection of Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL).
No significant cross-reactivity or interference between Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL) and the recovery rates were calculated by comparing the measured value to the expected amount of Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 82-90 86
EDTA plasma(n=5) 78-101 83
heparin plasma(n=5) 94-102 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 89-96% 98-105% 84-93% 78-98%
EDTA plasma(n=5) 81-102% 86-94% 94-104% 80-94%
heparin plasma(n=5) 85-93% 88-102% 86-96% 82-90%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
Substrate A 1×10mL Substrate B 1×2mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 100µL Substrate Solution. Incubate 10 minutes at 37°C;
8. Read RLU value immediately.

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Magazine Citations
Archives of Oral Biology Compressive stress induced the up-regulation of M-CSF, RANKL, TNF-α expression and the down-regulation of OPG expression in PDL cells via the integrin-FAK pathway ScienceDirect: S0003996912003998
PLOS One Adipokines in Psoriatic Arthritis Patients: The Correlations with Osteoclast Precursors and Bone Erosions PubMed: PMC3483160
Arthritis Research&Therapy Regulatory effect of calcineurin inhibitor, tacrolimus, on IL-6/sIL-6R-mediated RANKL expression through JAK2-STAT3-SOCS3 signaling pathway in fibroblast-like synoviocytes PubMed: PMC3672788
International Journal of Oral and Maxillofacial Surgery The effects of chronic zoledronate usage on the jaw and long bones evaluated using RANKL and osteoprotegerin levels in an animal model Pubmed: 23522850
AGE Estrogen deficiency does not decrease the in vitro osteogenic potential of rat adipose-derived mesenchymal stem cells Pubmed: 24687841
Food Science & Nutrition Effects and mechanisms of 8-prenylnaringenin on osteoblast MC3T3-E1 and osteoclast-like cells RAW264.7 Onlinelibrary: fsn3.109
Food Sci Nutr. Effects and mechanisms of 8‐prenylnaringenin on osteoblast MC3T3‐E1 and osteoclast‐like cells RAW264. 7 Pubmed:Pmc4221832
Journal of Investigative Surgery Effect of Nicotine on RANKL and OPG and Bone Mineral Density Pubmed:24830656
Int J Mol Med. Effects of icariin on the regulation of the OPG?RANKL?RANK system are mediated through the MAPK pathways in IL?1β?stimulated human SW1353 chondrosarcoma … Pubmed:25270538
Clinical and Experimental Pharmacology and Physiology Carvedilol promotes neurological function, reduces bone loss and attenuates cell damage after acute spinal cord injury in rats Pubmed:25424914
Int J Clin Exp Pathol A novel model of bisphosphonate-related osteonecrosis of the jaw in rats PubMed: 26191212
oncotarget MicroRNA-106b inhibits osteoclastogenesis and osteolysis by targeting RANKL in giant cell tumor of bone PubMed: 26053181
European Journal of Clinical Investigation Serum osteopontin levels are upregulated and predict disability after an ischaemic stroke PubMed: 25845543
Scienze Biomediche Effetti del laser a bassa dose su cellule osteoblastiche INCERTIPARENTI_SERENA_TESI.pdf
Nutrition. Noni leaf and black tea enhance bone regeneration in estrogen-deficient rats. pubmed:27908549
PLoS One. Extracorporeal shock waves alone or combined with raloxifene promote bone formation and suppress resorption in ovariectomized rats. pubmed:28158228
Epilepsy Research The effect of lamotrigine and phenytoin on bone turnover and bone strength: A prospective study in Wistar rats pubmed:27838501
Int Endod J.  Odontoclastogenesis of mouse papilla-derived MDPC-23 cells induced by lipopolysaccharide. pubmed:28333374
Sci Rep.  Sprint Interval Training Induces A Sexual Dimorphism but does not Improve Peak Bone Mass in Young and Healthy Mice. pubmed:28303909
Sci Rep. Skeletal Site-specific Effects of Zoledronate on in vivo Bone Remodeling and in vitro BMSCs Osteogenic Activity. pubmed:28139685
Biomedicine & Pharmacotherapy Anti-osteoporotic effects of an antidepressant tianeptine on ovariectomized rats S0753332216322752
BMJ Journals SAT0003 The Role of 12/15-Lipoxygenase in The Pathogenesis of Osteoporosis in Mice content:75
Japanese Society for Bone and Mineral Research Allogeneic yet major histocompatibility complex‑matched bone marrow transplantation in mice results in an impairment of osteoblasts and a significantly reduced trabecular bone article:10.1007
ONCOLOGY LETTERS Assessment of bone metabolism and biomechanical properties of the femur, following treatment with anastrozole and letrozole in an experimental model of menopause ol.2017.6596
Journal of Clinical Periodontology  Peroxisome proliferator‐activated receptor γ plays dual roles on experimental periodontitis in rats Pubmed:29574908
Journal of Functional Foods Effect of long-term administration of mangiferin from on bone metabolism in ovariectomized rats 10.1016:j.jff.2018.04.048
International Immunopharmacology Switched memory B cells promote alveolar bone damage during periodontitis: An adoptive transfer experiment Pubmed:30015235
European Journal of Pharmacology The effect of levetiracetam on rat bone mineral density, bone structure and biochemical markers of bone metabolism Pubmed:29428468
Journal of Bone and Mineral Metabolism Allogeneic yet major histocompatibility complex-matched bone marrow transplantation in mice results in an impairment of osteoblasts and a significantly reduced … Pubmed:28766138
Experimental and Therapeutic Medicine Liraglutide exerts a bone‑protective effect in ovariectomized rats with streptozotocin‑induced diabetes by inhibiting osteoclastogenesis Pubmed:29805533
Bone The sole and combined effect of simvastatin and platelet rich fibrin as a filling material in induced bone defect in tibia of albino rats Pubmed: 30208342
Journal of Periodontal Research Improved RANKL expression and osteoclastogenesis induction of CD27+ CD38− memory B cells: A link between B cells and alveolar bone damage in periodontitis Pubmed: 30346027
Bone Research NUMB maintains bone mass by promoting degradation of PTEN and GLI1 via ubiquitination in osteoblasts Pubmed: 30455992
Journal of International Dental and Medical Research Alendronate Effect in RANKL Concentration at Ovariectomized Sprague Dawley Rat.
Bone Development and characterization of supramolecular calcitonin assembly and assessment of its interactions with the bone remodelling process Pubmed: 30797058
Journal of Oral and Maxillofacial Surgery, Medicine, and Pathology Evaluation of therapeutic effects of teriparatide in a rat model of zoledronic acid-induced bisphosphonate-related osteonecrosis
Rheumatology International Comparison of RANKL expression, inflammatory markers, and cardiovascular risk in patients with acute coronary syndrome with and without rheumatoid arthritis Pubmed: 31297563
World Journal of Pediatrics Regulation of JAK/STAT signal pathway by miR-21 in the pathogenesis of juvenile idiopathic arthritis Pubmed: 31641939
Bone Mice lacking plastin-3 display a specific defect of cortical bone acquisition Pubmed: 31678489
BIOSCIENCE REPORTS IL-38 restrains inflammatory response of collagen-induced arthritis in rats via SIRT1/HIF-1α signaling pathway Pubmed: 32347300
Journal of Ethnopharmacology Anti-osteoporosis effect of Semen Cuscutae in ovariectomized mice through inhibition of bone resorption by osteoclasts 34801609
Br J Haematol Inducible T‐cell co‐stimulator (ICOS) and ICOS ligand are novel players in the multiple‐myeloma microenvironment Pubmed:34954822
Biomed Pharmacother Jiangu granule ameliorated OVX rats bone loss by modulating gut microbiota-SCFAs-Treg/Th17 axis Pubmed:35453007
Phytomedicine Transcriptomic profiling revealed the role of apigenin-4′-O-α-L-rhamnoside in inhibiting the activation of rheumatoid arthritis fibroblast-like synoviocytes via MAPK … Pubmed:35660352
Med Science Identification of marker proteins in preterm and term birth: Relationship of rankl and HSP70 in the cord blood
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