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|Antigen||Transforming Growth Factor, beta 1 (TGFB1) ELISA Kits|
|Reactivity||Mouse (Murine) Alternatives|
Kits with alternative reactivity to:
|Method Type||Sandwich ELISA|
|Detection Range||15.6-1000 pg/mL|
|Minimum Detection Limit||15.6 pg/mL|
|9 references available|
|Supplier||Log in to see|
Product Details TGFB1 ELISA KitTarget details Application Details Handling ProductDetails: References for TGFB1 Kit (ABIN415536) Images
|Purpose||The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of TGFb1 in Serum,Platelet-Poor Plasma,Tissue Homogenate,Cell Culture Supernatant,Biological Fluids|
|Sample Type||Serum, Platelet-Poor Plasma, Tissue Homogenate, Cell Culture Supernatant, Biological Fluids|
This assay has high sensitivity and excellent specificity for detection of Transforming Growth Factor Beta 1 (TGFb1).
|Cross-Reactivity (Details)||No significant cross-reactivity or interference between Transforming Growth Factor Beta 1 (TGFb1) and analogues was observed.|
|Material not included||
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|Alternative Name||TGFb1 (TGFB1 ELISA Kit Abstract)|
|Research Area||Growth Factors, Signaling|
|Pathways||EGFR Signaling Pathway, Dopaminergic Neurogenesis, Cellular Response to Molecule of Bacterial Origin, Glycosaminoglycan Metabolic Process, Regulation of Leukocyte Mediated Immunity, Regulation of Muscle Cell Differentiation, Positive Regulation of Immune Effector Process, Cell-Cell Junction Organization, Production of Molecular Mediator of Immune Response, Ribonucleoside Biosynthetic Process, Skeletal Muscle Fiber Development, Regulation of Carbohydrate Metabolic Process, Protein targeting to Nucleus, Autophagy|
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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 Transforming Growth Factor Beta 1 (TGFb1). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Transforming Growth Factor Beta 1 (TGFb1). 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 Transforming Growth Factor Beta 1 (TGFb1), 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 Transforming Growth Factor Beta 1 (TGFb1) 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
Platelet-Poor Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 × g at 2-8°C within 30 minutes of collection. It is recommended to centrifuge samples for 10 minutes at 10,000 × g for complete platelet removal. Remove plasma and assay (see activation procedure) immediately or store samples in aliquot at -20°C or -80°C for later use. Avoid repeated freeze/thaw cycles.
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 TGFb1 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 Transforming Growth Factor Beta 1 (TGFb1) were tested 20 times on one plate, respectively.
|Restrictions||For Research Use only|
HandlingProduct Details TGFB1 ELISA Kit Target details Application Details ProductDetails: References for TGFB1 Kit (ABIN415536) 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.
|Expiry Date||6 months|
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|Product cited in:||
Kabel, Abd Elmaaboud, Atef, Baali: "Ameliorative potential of linagliptin and/or calcipotriol on bleomycin-induced lung fibrosis: In vivo and in vitro study." in: Environmental toxicology and pharmacology, Vol. 50, pp. 216-226, 2017
He, Huang, Chen, Chen, Zhang, Li, Chen, Chen: "Recombinant Mip-PilE-FlaA dominant epitopes vaccine candidate against Legionella pneumophila." in: Immunology letters, Vol. 186, pp. 33-40, 2017
Xia, Chang, Zhang, Shi, Liu, Ding, Liu, Gao, Dong: "Therapeutic effects of bone marrow-derived mesenchymal stem cells on radiation-induced lung injury." in: Oncology reports, Vol. 35, Issue 2, pp. 731-8, 2016
Wang, Zhang, Sun, Ren: "The protective role of vitamin D3 in a murine model of asthma via the suppression of TGF-β/Smad signaling and activation of the Nrf2/HO-1 pathway." in: Molecular medicine reports, Vol. 14, Issue 3, pp. 2389-96, 2016
Zhang, Ju, Zhang, Zhu, Nie, Xu, Lei et al.: "Magnolol Attenuates Concanavalin A-induced Hepatic Fibrosis, Inhibits CD4(+) T Helper 17 (Th17) Cell Differentiation and Suppresses Hepatic Stellate Cell Activation: Blockade of Smad3/Smad4 ..." in: Basic & clinical pharmacology & toxicology, Vol. 120, Issue 6, pp. 560-570, 2016
Wang, Liu, Bian, Sun, Du, Wang, Su, Li: "Epigallocatechin-3-gallate reduces tubular cell apoptosis in mice with ureteral obstruction." in: The Journal of surgical research, Vol. 197, Issue 1, pp. 145-54, 2015
Yu, Sun, Feng, Tan, Fang, Zhao, Zhao, Pu, Huang, Xiang, Cao, He: "MSX3 Switches Microglia Polarization and Protects from Inflammation-Induced Demyelination." in: The Journal of neuroscience : the official journal of the Society for Neuroscience, Vol. 35, Issue 16, pp. 6350-65, 2015
Obrador, Benlloch, Pellicer, Asensi, Estrela: "Intertissue flow of glutathione (GSH) as a tumor growth-promoting mechanism: interleukin 6 induces GSH release from hepatocytes in metastatic B16 melanoma-bearing mice." in: The Journal of biological chemistry, Vol. 286, Issue 18, pp. 15716-27, 2011 Method employed by authors: ELISA (ELISA) (Sample species: Mouse (Murine)).
Xiao, Ma, Feng, Fu, Lu, Xu, Shen, Zhu, Zhang: "Metformin attenuates cardiac fibrosis by inhibiting the TGFbeta1-Smad3 signalling pathway." in: Cardiovascular research, Vol. 87, Issue 3, pp. 504-13, 2010
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