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|Application / Reactivity||Mouse (Murine)|
|ELISA||9 ELISA Kits|
|Antigen||High-Mobility Group Box 1 (HMGB1) ELISA Kits|
|Reactivity||Mouse (Murine) Alternatives|
Kits with alternative reactivity to:
|Methode Type||Sandwich ELISA|
|Detection Range||46.88-3000 pg/mL|
|Minimum Detection Limit||46.88 pg/mL|
|10 references available|
|Supplier||Log in to see|
Product Details HMGB1 ELISA KitTarget details Application Details Handling References for HMGB1 Kit (ABIN415379) Images
|Purpose||The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of HMG1 in Serum,Plasma,Biological Fluids|
|Sample Type||Serum, Plasma, Biological Fluids|
|Specificity||This assay has high sensitivity and excellent specificity for detection of High Mobility Group Protein 1 (HMG1).|
|Cross-Reactivity (Details)||No significant cross-reactivity or interference between High Mobility Group Protein 1 (HMG1) 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||HMG1 (HMGB1 ELISA Kit Abstract)|
Application DetailsProduct Details HMGB1 ELISA Kit Target details Handling References for HMGB1 Kit (ABIN415379) 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 HMG1. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific for HMG1.
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 HMG1, 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 HMG1 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.
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 HMG1 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 mouse HMG1 were tested 20 times on one plate, respectively.
Inter-assay Precision (precision between assays): 3 samples with low, middle and high level mouse HMG1 were tested on 3 different plates, 8 replicates in each plate.
CV (%) = SD/mean X 100
|Restrictions||For Research Use only|
HandlingProduct Details HMGB1 ELISA Kit Target details Application Details References for HMGB1 Kit (ABIN415379) 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 HMGB1 Kit (ABIN415379)Product Details HMGB1 ELISA Kit Target details Application Details Handling Images back to top
|Product cited in:||
Kim, Ha, Kim, Park, Kim, Park, Kim, Chung, Chang: "Ascorbic acid reduces HMGB1 secretion in lipopolysaccharide-activated RAW 264.7 cells and improves survival rate in septic mice by activation of Nrf2/HO-1 signals." in: Biochemical pharmacology, Vol. 95, Issue 4, pp. 279-89, 2015
Sun, Chen, Dai, Zou, Gao, Wu, Ming, Lai, Xiao, Xiong, Xu, Gong, Zheng: "HMGB1 expression patterns during the progression of experimental autoimmune encephalomyelitis." in: Journal of neuroimmunology, Vol. 280, pp. 29-35, 2015
Kim, Kim, Chang: "Glycyrrhizin reduces HMGB1 secretion in lipopolysaccharide-activated RAW 264.7 cells and endotoxemic mice by p38/Nrf2-dependent induction of HO-1." in: International immunopharmacology, Vol. 26, Issue 1, pp. 112-8, 2015
Chen, Sun, Lai, Wu, Xiao, Ming, Gao, Zou, Xiong, Xu, Tan, Gong, Zheng: "Interleukin-33 is released in spinal cord and suppresses experimental autoimmune encephalomyelitis in mice." in: Neuroscience, Vol. 308, pp. 157-68, 2015
Jiang, Wang, Yun, Hajrasouliha, Zhao, Sun, Kaplan, Shao: "HMGB1 release triggered by the interaction of live retinal cells and uveitogenic T cells is Fas/FasL activation-dependent." in: Journal of neuroinflammation, Vol. 12, pp. 179, 2015
Kim, Kim, Park, Kim, Chang: "Retinoic acid inhibits tissue factor and HMGB1 via modulation of AMPK activity in TNF-? activated endothelial cells and LPS-injected mice." in: Atherosclerosis, Vol. 241, Issue 2, pp. 615-23, 2015
Jiang, Sun, Yang, Lu, Kaplan, Shao: "HMGB1 is an early and critical mediator in an animal model of uveitis induced by IRBP-specific T cells." in: Journal of leukocyte biology, Vol. 95, Issue 4, pp. 599-607, 2014
Liu, Yu, Jiang, Zhang, Zhang, Yu, Jia, Chen, Zou, Ge: "Simvastatin suppresses vascular inflammation and atherosclerosis in ApoE(-/-) mice by downregulating the HMGB1-RAGE axis." in: Acta pharmacologica Sinica, Vol. 34, Issue 6, pp. 830-6, 2013
Wang, Zhang, Lei, Zhou, Ye: "The inhibitory effect of lidocaine on the release of high mobility group box 1 in lipopolysaccharide-stimulated macrophages." in: Anesthesia and analgesia, Vol. 112, Issue 4, pp. 839-44, 2011
Ni, Tian, Lu, Yang, Fu, Wang, Yan, Zhao, Wang, Jiang: "Protective effect of nicotine on lipopolysaccharide-induced acute lung injury in mice." in: Respiration; international review of thoracic diseases, Vol. 81, Issue 1, pp. 39-46, 2010
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