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|Application / Reactivity||Mouse (Murine)|
|ELISA||15 ELISA Kits|
|Antigen||Vascular Endothelial Growth Factor A (VEGFA) ELISA Kits|
|Reactivity||Mouse (Murine) Alternatives|
Kits with alternative reactivity to:
|Methode Type||Sandwich ELISA|
|Detection Range||15.63-1000 pg/mL|
|Minimum Detection Limit||15.63 pg/mL|
|10 references available|
|Supplier||Log in to see|
Product Details VEGF ELISA KitTarget details Application Details Handling References for VEGF Kit (ABIN424307) Images
|Purpose||The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of VEGFA 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 Vascular Endothelial Growth Factor A (VEGFA).|
|Cross-Reactivity (Details)||No significant cross-reactivity or interference between Vascular Endothelial Growth Factor A (VEGFA) 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 VEGF ELISA Kit Application Details Handling References for VEGF Kit (ABIN424307) Images back to top
|Alternative Name||VEGFA (VEGFA ELISA Kit Abstract)|
|Research Area||Cancer, Growth Factors, Angiogenesis|
Application DetailsProduct Details VEGF ELISA Kit Target details Handling References for VEGF Kit (ABIN424307) 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 VEGFA. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific for VEGFA.
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 VEGFA, 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 VEGFA 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 VEGFA 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 mouse VEGFA were tested 20 times on one plate, respectively.
Inter-assay Precision (precision between assays): 3 samples with low, middle and high level mouse VEGFA were tested on 3 different plates, 8 replicates in each plate.
CV (%) = SD/mean X 100
|Restrictions||For Research Use only|
HandlingProduct Details VEGF ELISA Kit Target details Application Details References for VEGF Kit (ABIN424307) 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 VEGF Kit (ABIN424307)Product Details VEGF ELISA Kit Target details Application Details Handling Images back to top
|Product cited in:||
Jiang, Zhang, Zhang, Xia: "Application of recombinant peroxisome proliferator-activated receptor-γ coactivator-1α mediates neovascularization in the retina." in: Molecular medicine reports, Vol. 13, Issue 2, pp. 1311-9, 2016
Gao, Guo, Zeng, Wang, Yang, Pang, Li, Suo, Jiang, Yu: "Matrix metalloproteinase 9 secreted by hypoxia cardiac fibroblasts triggers cardiac stem cell migration in vitro." in: Stem cells international, Vol. 2015, pp. 836390, 2015
He, Hu, Yan, Li, Zhang, Zhang, Chen, Huang: "Pigment epithelium-derived factor regulates microvascular permeability through adipose triglyceride lipase in sepsis." in: Clinical science (London, England : 1979), Vol. 129, Issue 1, pp. 49-61, 2015
Peng, Gao, Jiang, Wang, Zhang, Xiao, Ou, Yang, Yang, Xiao: "ANGIOGENESIS INHIBITOR ENDOSTATIN PROTECTS MICE WITH SEPSIS FROM MULTIPLE ORGAN DYSFUNCTION SYNDROME." in: Shock (Augusta, Ga.), Vol. 44, Issue 4, pp. 357-64, 2015
Cai, Li, Wang, Shi, Ye, Yang, Dou, Hou, Yang, Cao, Lu: "Hyperglycemia promotes vasculogenesis in choroidal neovascularization in diabetic mice by stimulating VEGF and SDF-1 expression in retinal pigment epithelial cells." in: Experimental eye research, Vol. 123, pp. 87-96, 2014
Wang, Chen, Yang, Zeng, Qi, Guo: "A bone-seeking clone exhibits different biological properties from the ACHN parental human renal cell carcinoma in vivo and in vitro." in: Oncology reports, Vol. 27, Issue 4, pp. 1104-10, 2012
Wang, Xu, Man, Zhang, Chu, Chu, Cheng, Li, He, Qin, Lau, Kwong, Chan: "Prodrug of green tea epigallocatechin-3-gallate (Pro-EGCG) as a potent anti-angiogenesis agent for endometriosis in mice." in: Angiogenesis, Vol. 16, Issue 1, pp. 59-69, 2012
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)).
Ono, Hirano: "Effects of ethanol on the riboflavin metabolism in the lens." in: International journal for vitamin and nutrition research. Internationale Zeitschrift für Vitamin- und Ernährungsforschung. Journal international de vitaminologie et de nutrition, Vol. 57, Issue 1, pp. 106, 1987
Goldblatt: "Whatever happened to the Black report?" in: British medical journal (Clinical research ed.), Vol. 293, Issue 6545, pp. 503-4, 1986
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