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|Application / Reactivity||Human|
|ELISA||13 ELISA Kits|
|Antigen||Interferon, beta 1, Fibroblast (IFNB1) ELISA Kits|
Kits with alternative reactivity to:
|Methode Type||Sandwich ELISA|
|Detection Range||7.81-500 pg/mL|
|Minimum Detection Limit||7.81 pg/mL|
|9 references available|
|Supplier||Log in to see|
Product Details IFNB1 ELISA KitTarget details Application Details Handling References for IFNB1 Kit (ABIN414734) Images
|Purpose||The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of IFNb 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 Interferon Beta (IFNb).|
|Cross-Reactivity (Details)||No significant cross-reactivity or interference between Interferon Beta (IFNb) 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||
Target detailsProduct Details IFNB1 ELISA Kit Application Details Handling References for IFNB1 Kit (ABIN414734) Images back to top
|Alternative Name||IFNb (IFNB1 ELISA Kit Abstract)|
Application DetailsProduct Details IFNB1 ELISA Kit Target details Handling References for IFNB1 Kit (ABIN414734) Images back to top
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 IFNb. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to IFNb.
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 IFNb, 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 IFNb 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. 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 IFNb 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 IFNb were tested 20 times on one plate, respectively.
Inter-assay Precision (precision between assays): 3 samples with low, middle and high level IFNb were tested on 3 different plates, 8 replicates in each plate.
CV (%) = SD/mean X 100
|Restrictions||For Research Use only|
HandlingProduct Details IFNB1 ELISA Kit Target details Application Details References for IFNB1 Kit (ABIN414734) 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 IFNB1 Kit (ABIN414734)Product Details IFNB1 ELISA Kit Target details Application Details Handling Images back to top
|Product cited in:||
Xie, Liu, Wu, Lin, Ma, Xiong, Wang, Li, Ma, Tu: "Effects of IRF1 and IFN-β interaction on the M1 polarization of macrophages and its antitumor function." in: International journal of molecular medicine, Vol. 38, Issue 1, pp. 148-60, 2016
Yang, Qin, Zhang, Yang, Xiang, Wei: "Cellular uptake of exogenous calcineurin B is dependent on TLR4/MD2/CD14 complexes, and CnB is an endogenous ligand of TLR4." in: Scientific reports, Vol. 6, pp. 24346, 2016
Liu, Qin, Kong, Shao, Su, Wang, Chen: "siRNA Targeting the 2Apro Genomic Region Prevents Enterovirus 71 Replication In Vitro." in: PLoS ONE, Vol. 11, Issue 2, pp. e0149470, 2016
Pinegin, Pashenkov, Kulakov, Murugin, Zhmak: "Complexes of DNA with the Antimicrobial Peptide LL37 Augment NK Cell Functions by Inducing Type I Interferon Production from Circulating Monocytes and Plasmacytoid Predendritic Cells." in: Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research, Vol. 35, Issue 11, pp. 850-8, 2015
Frellstedt, Waldschmidt, Gosset, Desmet, Pirottin, Bureau, Farnir, Franck, Dupuis-Tricaud, Lekeux, Art: "Training modifies innate immune responses in blood monocytes and in pulmonary alveolar macrophages." in: American journal of respiratory cell and molecular biology, Vol. 51, Issue 1, pp. 135-42, 2014
Liu, Zhou, Ma, Lu, Ming, Shan, Zhang, Hou, Chen, Zuo: "Mannan binding lectin attenuates double-stranded RNA-mediated TLR3 activation and innate immunity." in: FEBS letters, Vol. 588, Issue 6, pp. 866-72, 2014
Zhu, He, Hu, Wen, Lin, Yu, Pan, Li, Deng, Liao, Yuan, Wu, Li, Li: "MicroRNA-30e* suppresses dengue virus replication by promoting NF-?B-dependent IFN production." in: PLoS neglected tropical diseases, Vol. 8, Issue 8, pp. e3088, 2014
Yu, Chen, Wu, Chen, Kato, Yuan et al.: "Hepatitis B virus polymerase inhibits RIG-I- and Toll-like receptor 3-mediated beta interferon induction in human hepatocytes through interference with interferon regulatory factor 3 activation and ..." in: The Journal of general virology, Vol. 91, Issue Pt 8, pp. 2080-90, 2010
Li, Li, Qian, Zhang, Chen, Shi: "Impaired TLR3/IFN-beta signaling in monocyte-derived dendritic cells from patients with acute-on-chronic hepatitis B liver failure: relevance to the severity of liver damage." in: Biochemical and biophysical research communications, Vol. 390, Issue 3, pp. 630-5, 2009
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