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|Application / Reactivity||Rat (Rattus)|
|ELISA||17 ELISA Kits|
|Antigen||Interleukin 17A (IL17A) ELISA Kits|
|Reactivity||Rat (Rattus) Alternatives|
Kits with alternative reactivity to:
|Methode Type||Sandwich ELISA|
|Detection Range||15.63-1000 pg/mL|
|Minimum Detection Limit||15.63 pg/mL|
|11 references available|
|Supplier||Log in to see|
Product Details IL17A ELISA KitTarget details Application Details Handling References for IL17A Kit (ABIN416366) Images
|Purpose||The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of IL17 in Serum,Plasma,Tissue Homogenate,Cell Lysate,Cell Culture Supernatant,Biological Fluids|
|Sample Type||Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids|
|Specificity||This assay has high sensitivity and excellent specificity for detection of Interleukin 17 (IL17).|
|Cross-Reactivity (Details)||No significant cross-reactivity or interference between Interleukin 17 (IL17) 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.
|Material not included||
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|Alternative Name||IL17 (IL17A ELISA Kit Abstract)|
|Research Area||Cancer, Cytokines, Innate Immunity|
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Information on standard material:
|Sample Volume||100 μL|
|Assay Time||4.5 h|
|Plate||Pre-coated,Strips (12 x 8)|
The microtiter plate provided in this kit has been pre-coated with an antibody specific to IL17. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific for IL17.
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 IL17, 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 450 nm ± 10 nm. The concentration of IL17 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
Cell Lysate: Cells must be lysed before assaying according to the following directions. Adherent cells should be detached with trypsin and then collected by centrifugation (suspension cells can be collected by centrifugation directly). Wash cells three times in cold PBS. Resuspend cells in PBS (1×) and subject them to ultrasonication for 4 times (or Freeze cells at -20 °C. Thaw cells with gentle mixing. Repeat the freeze/thaw cycle for 3 times.) Centrifuge at 1500 × g for 10 minutes at 2 - 8°C to remove cellular debris.
Cell Culture Supernatant: 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.
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 IL17 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 IL17 were tested 20 times on one plate, respectively.
Inter-assay Precision (precision between assays): 3 samples with low, middle and high level rat IL17 were tested on 3 different plates, 8 replicates in each plate.
CV (%) = SD/mean X 100
|Restrictions||For Research Use only|
HandlingProduct Details IL17A ELISA Kit Target details Application Details References for IL17A Kit (ABIN416366) 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||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 IL17A Kit (ABIN416366)Product Details IL17A ELISA Kit Target details Application Details Handling Images back to top
|Product cited in:||
Liu, Wang, Wei, Fan, Lu, Zhu, Chen, Huang, Zhou, Zheng: "Consistency and pathophysiological characterization of a rat polymicrobial sepsis model via the improved cecal ligation and puncture surgery." in: International immunopharmacology, Vol. 32, pp. 66-75, 2016
Yan, Yang, Han, Feng: "Tanshinone IIA attenuates experimental autoimmune encephalomyelitis in rats." in: Molecular medicine reports, Vol. 14, Issue 2, pp. 1601-9, 2016
Karatas, Koca, Ozgen, Dagli, Erman, Sahin, Sahin, Isik: "Pemetrexed ameliorates experimental arthritis in rats." in: Inflammation, Vol. 38, Issue 1, pp. 9-15, 2015
Ozgen, Koca, Karatas, Dagli, Erman, Gundogdu, Sahin, Isik: "Lapatinib ameliorates experimental arthritis in rats." in: Inflammation, Vol. 38, Issue 1, pp. 252-9, 2015
Zhu, Wang, Sun, Wu: "Protective effect and mechanism of probucol in the treatment of spinal cord injury." in: Genetics and molecular research : GMR, Vol. 14, Issue 3, pp. 8029-37, 2015
Xu, Tan, Hu, Alwahsh, Yan, Hu, Dai, Wang, Zhou, Fan, Huang: "Expression of hemopexin in acute rejection of rat liver allograft identified by serum proteomic analysis." in: Shock (Augusta, Ga.), Vol. 42, Issue 1, pp. 65-74, 2014
Nasrin, Masuda, Kugaya, Ito, Yamada: "Improvement by phytotherapeutic agent of detrusor overactivity, down-regulation of pharmacological receptors and urinary cytokines in rats with cyclophosphamide induced cystitis." in: The Journal of urology, Vol. 189, Issue 3, pp. 1123-9, 2013
Li, Dou, Liu, Shi, Cao, Zhang, Zhu, Duan: "Atorvastatin ameliorates experimental autoimmune neuritis by decreased Th1/Th17 cytokines and up-regulated T regulatory cells." in: Cellular immunology, Vol. 271, Issue 2, pp. 455-61, 2011
Andersson, Isaksson, Wefer, Norling, Flores-Morales, Rorsman, Kämpe, Harris, Lobell: "Impaired autoimmune T helper 17 cell responses following DNA vaccination against rat experimental autoimmune encephalomyelitis." in: PLoS ONE, Vol. 3, Issue 11, pp. e3682, 2008
Johnson, Drugan, Miller, Evans: "Genetic correction of hereditary disease." in: Fetal therapy, Vol. 4 Suppl 1, pp. 28-39, 1991
Johnson, Drugan, Miller, Evans: "38" in: , Vol. 1363, Issue Nucleic acids research, pp. 28-39, 1991
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