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|Antigen||Tumor Necrosis Factor (Ligand) Superfamily, Member 11 (TNFSF11) ELISA Kits|
|Reactivity||Rat (Rattus) Alternatives|
Kits with alternative reactivity to:
|Method Type||Sandwich ELISA|
|Detection Range||15.6 pg/mL - 1000 pg/mL|
|Minimum Detection Limit||15.6 pg/mL|
|8 references available|
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Product Details TNFSF11 ELISA KitTarget Details Application Details Handling References for TNFSF11 Kit (ABIN416407) Images
|Purpose||The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of RANkL in Serum,Plasma,Tissue Homogenate,Biological Fluids|
|Sample Type||Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate|
This assay has high sensitivity and excellent specificity for detection of Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL).
|Cross-Reactivity (Details)||No significant cross-reactivity or interference between Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL) and analogues was observed.|
|Material not included||
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|Alternative Name||RANkL (TNFSF11 ELISA Kit Abstract)|
Application DetailsProduct Details TNFSF11 ELISA Kit Target Details Handling References for TNFSF11 Kit (ABIN416407) Images back to top
Information on standard material:
|Sample Volume||100 μL|
|Assay Time||3 h|
|Protocol||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 Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL). 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 Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL), 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 Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL) in the samples is then determined by comparing the O.D. of the samples to the standard curve.|
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.
Tissue Homogenates: The preparation of tissue homogenates will vary depending upon tissue type. For this assay, rinse tissues in ice-cold PBS (0.02mol/L,pH 7.0-7.2) to remove excess blood thoroughly and weigh before homogenization. Mince the tissues to small pieces and homogenize them in 5-10 mL of PBS with a glass homogenizer on ice (Micro Tissue Grinders work, too). Sonicate the resulting suspension with an ultrasonic cell disrupter or subject it to two freeze-thaw cycles to further break the cell membranes. Centrifugate the homogenates for 5 minutes at 5000 × g. Remove the supernate and assay immediately or aliquot and store at -20°C
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. Construct a standard curve by plotting the mean O.D. and concentration for each standard and draw a best fit curve through the points on the graph or create a standard curve on log-log graph paper with RANkL concentration on the y-axis and absorbance on the x-axis. 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 Receptor Activator Of Nuclear Factor Kappa B Ligand (RANkL) were tested 20 times on one plate, respectively.
|Restrictions||For Research Use only|
HandlingProduct Details TNFSF11 ELISA Kit Target Details Application Details References for TNFSF11 Kit (ABIN416407) 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.|
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.
|Expiry Date||6 months|
References for TNFSF11 Kit (ABIN416407)Product Details TNFSF11 ELISA Kit Target Details Application Details Handling Images back to top
|Product cited in:||
Gong, Yu, Zhao, Su, Sheng: "Skeletal Site-specific Effects of Zoledronate on in vivo Bone Remodeling and in vitro BMSCs Osteogenic Activity." in: Scientific reports, Vol. 7, pp. 36129, 2017
Lama, Santoro, Corrado, Pirozzi, Paciello, Pagano, Russo, Calignano, Mattace Raso, Meli: "Extracorporeal shock waves alone or combined with raloxifene promote bone formation and suppress resorption in ovariectomized rats." in: PLoS ONE, Vol. 12, Issue 2, pp. e0171276, 2017
Simko, Karesova, Kremlacek, Fekete, Zimcikova, Malakova, Zivna, Valis, Palicka: "The effect of lamotrigine and phenytoin on bone turnover and bone strength: A prospective study in Wistar rats." in: Epilepsy research, Vol. 128, pp. 113-118, 2016
Shalan, Mustapha, Mohamed: "Noni leaf and black tea enhance bone regeneration in estrogen-deficient rats." in: Nutrition (Burbank, Los Angeles County, Calif.), Vol. 33, pp. 42-51, 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
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
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
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