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|Application / Reactivity||Rat (Rattus)|
|ELISA||14 ELISA Kits|
|Antigen||Tumor Necrosis Factor (Ligand) Superfamily, Member 11 (TNFSF11) ELISA Kits|
|Reactivity||Rat (Rattus) Alternatives|
Kits with alternative reactivity to:
|Methode Type||Sandwich ELISA|
|Detection Range||0.15-10 ng/mL|
|Minimum Detection Limit||0.15 ng/mL|
|11 references available|
|Supplier||Log in to see|
Product Details TNFSF11 ELISA KitTarget details Application Details Handling References for TNFSF11 Kit (ABIN416342) Images
|Purpose||The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of OPG in Serum,Plasma,Biological Fluids|
|Sample Type||Serum, Plasma, Tissue Homogenate, Biological Fluids|
|Specificity||This assay has high sensitivity and excellent specificity for detection of Osteoprotegerin (OPG).|
|Cross-Reactivity (Details)||No significant cross-reactivity or interference between Osteoprotegerin (OPG) and analogues was observed.|
The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero. It was determined by adding two standard deviations - the mean optical density value of twenty zero standard replicates and calculating the corresponding concentration.
|Characteristics||The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Osteoprotegerin (OPG). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Osteoprotegerin (OPG). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Osteoprotegerin (OPG), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Osteoprotegerin (OPG) in the samples is then determined by comparing the O.D. of the samples to the standard curve.|
Pre-coated, ready to use 96-well strip plate: 1
Plate sealer for 96 wells: 4
Standard Diluent: 1×20 mL
Detection Reagent A: 1×120 µL
Assay Diluent: A 1×12 mL
Detection Reagent B: 1×120 µL
Assay Diluent B: 1×12 mL
Substrate A: 1×10 mL
Substrate B: 1×2 mL
Wash Buffer (30 × concentrate): 1×20 mL
Instruction manual: 1
|Material not included||
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|Alternative Name||OPG (TNFSF11 ELISA Kit Abstract)|
Application DetailsProduct Details TNFSF11 ELISA Kit Target details Handling References for TNFSF11 Kit (ABIN416342) Images back to top
Information on standard material:
|Sample Volume||100 μL|
|Assay Time||4.5 h|
|Plate||Pre-coated,Strips (12 x 8),96 wells|
1. Prepare all reagents, samples and standards
2. Add 100 µL standard or sample to each well. Incubate 2 hours at 37 °C
3. Aspirate and add 100 µL prepared Detection Reagent A. Incubate 1 hour at 37 °C
4. Aspirate and wash 3 times
5. Add 100 µL prepared Detection Reagent B. Incubate 30 minutes at 37 °C
6. Aspirate and wash 5 times
7. Add 90 µL Substrate Solution. Incubate 15-25 minutes at 37 °C
8. Add 50 µL Stop Solution. Read at 450 nm immediately.
Serum: Allow samples to clot for two hours at room temperature or overnight at 4°C before centrifugation for 20 minutes at approximately 1000 × g. Assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 × g within 30 minutes of collection. Remove plasma and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
Biological Fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20 °C or -80 °C for later use. Avoid repeated freeze/thaw cycles.
|Calculation of Results||
Average the duplicate readings for each standard, control, and samples and subtract the average zero standard optical density. Create a standard curve on log-log graph paper, with OPG concentration on the y-axis and absorbance on the x-axis. Draw the best fit straight line through the standard points and it can be determined by regression analysis. Using some plot software, for instance, curve expert 1.30, is also recommended. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.
In order to make the calculation easier, we plot the O.D. value of the standard (X-axis) against the known concentration of the standard (Y-axis), although concentration is the independent variable and O.D. value is the dependent variable. However, the O.D. values of the standard curve may vary according to the conditions of assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), plotting log of the data to establish standard curve for each test is recommended. Typical standard curve below is provided for reference only.
Intra-assay Precision (precision within an assay): 3 samples with low, middle and high level rat OPG were tested 20 times on one plate, respectively.
Inter-assay Precision (precision between assays): 3 samples with low, middle and high level rat OPG were tested on 3 different plates, 8 replicates in each plate.
CV (%) = SD/mean X 100
|Restrictions||For Research Use only|
HandlingProduct Details TNFSF11 ELISA Kit Target details Application Details References for TNFSF11 Kit (ABIN416342) Images back to top
|Precaution of Use||The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.|
|Handling Advice||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.|
|Storage||4 °C/-20 °C|
|Expiry Date||6 months|
References for TNFSF11 Kit (ABIN416342)Product Details TNFSF11 ELISA Kit Target details Application Details Handling Images back to top
|Product cited in:||
Stringhetta-Garcia, Singulani, Santos, Louzada, Nakamune, Chaves-Neto, Rossi, Ervolino, Dornelles: "The effects of strength training and raloxifene on bone health in aging ovariectomized rats." in: Bone, Vol. 85, pp. 45-54, 2016
Chen, Wang, Wang, Yin, Gao, Xiang, Liu, Xiong, Wang, Zhu, Yang, Zhang: "Antiosteoporotic effect of icariin in ovariectomized rats is mediated via the Wnt/β-catenin pathway." in: Experimental and therapeutic medicine, Vol. 12, Issue 1, pp. 279-287, 2016
Liu, Huang, Li, Jia, Liang, Fu: "Carvedilol promotes neurological function, reduces bone loss and attenuates cell damage after acute spinal cord injury in rats." in: Clinical and experimental pharmacology & physiology, Vol. 42, Issue 2, pp. 202-12, 2015
Fekete, Simko, Mzik, Karesova, Zivna, Pavlíková, Palicka: "Effect of mirtazapine on rat bone tissue after orchidectomy." in: Pharmacology, Vol. 95, Issue 3-4, pp. 166-72, 2015
Yang, Pan, Yu, Chen, Shang, Xu: "A novel model of bisphosphonate-related osteonecrosis of the jaw in rats." in: International journal of clinical and experimental pathology, Vol. 8, Issue 5, pp. 5161-7, 2015
Veronesi, Pagani, Della Bella, Giavaresi, Fini: "Estrogen deficiency does not decrease the in vitro osteogenic potential of rat adipose-derived mesenchymal stem cells." in: Age (Dordrecht, Netherlands), Vol. 36, Issue 3, pp. 9647, 2014
Simko, Fekete, Gradosova, Malakova, Zivna, Valis, Palicka, Zivny: "The effect of topiramate and lamotrigine on rat bone mass, structure and metabolism." in: Journal of the neurological sciences, Vol. 340, Issue 1-2, pp. 80-5, 2014
Mizrak, Turan, Inan, Uysal, Yilmaz, Ercan: "Effect of nicotine on RANKL and OPG and bone mineral density." in: Journal of investigative surgery : the official journal of the Academy of Surgical Research, Vol. 27, Issue 6, pp. 327-31, 2014
Çankaya, Cizmeci ?enel, Kadioglu Duman, Muci, Dayisoylu, Balaban: "The effects of chronic zoledronate usage on the jaw and long bones evaluated using RANKL and osteoprotegerin levels in an animal model." in: International journal of oral and maxillofacial surgery, Vol. 42, Issue 9, pp. 1134-9, 2013
Gradosova, Zivna, Palicka, Hubena, Svejkovska, Zivny: "Protective effect of amlodipine on rat bone tissue after orchidectomy." in: Pharmacology, Vol. 89, Issue 1-2, pp. 37-43, 2012
Gradosova, Zivna, Palicka, Hubena, Svejkovska, Zivny: "Protective effect of atorvastatin on bone tissue in orchidectomised male albino Wistar rats." in: European journal of pharmacology, Vol. 679, Issue 1-3, pp. 144-50, 2012
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