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|Antigen||Vascular Endothelial Growth Factor A (VEGFA) ELISA Kits|
Kits with alternative reactivity to:
|Method Type||Sandwich ELISA|
|Detection Range||15.6-1000 pg/mL|
|Minimum Detection Limit||15.6 pg/mL|
|19 references available|
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Product Details VEGF ELISA KitTarget details Application Details Handling ProductDetails: References for VEGF Kit (ABIN414845) 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|
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.|
|Material not included||
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|Alternative Name||VEGFA (VEGFA ELISA Kit Abstract)|
|Research Area||Cancer, Growth Factors, Angiogenesis|
|Pathways||RTK Signaling, Glycosaminoglycan Metabolic Process, Regulation of Cell Size, Tube Formation, Signaling Events mediated by VEGFR1 and VEGFR2, Platelet-derived growth Factor Receptor Signaling, VEGFR1 Specific Signals|
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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 Vascular Endothelial Growth Factor A (VEGFA). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Vascular Endothelial Growth Factor A (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 Vascular Endothelial Growth Factor A (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 450nm ± 10nm. The concentration of Vascular Endothelial Growth Factor A (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 Vascular Endothelial Growth Factor A (VEGFA) were tested 20 times on one plate, respectively.
|Restrictions||For Research Use only|
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|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:||
Pagani, Torricelli, Veronesi, Salamanna, Cepollaro, Fini: "An advanced tri-culture model to evaluate the dynamic interplay among osteoblasts, osteoclasts, and endothelial cells." in: Journal of cellular physiology, 2017
Vurusaner, Gamba, Gargiulo, Testa, Staurenghi, Leonarduzzi, Poli, Basaga: "Nrf2 antioxidant defense is involved in survival signaling elicited by 27-hydroxycholesterol in human promonocytic cells." in: Free radical biology & medicine, Vol. 91, pp. 93-104, 2016
Biberoglu, Kirbas, Daglar, Biberoglu, Timur, Demirtas, Karabulut, Danisman: "Serum angiogenic profile in abnormal placentation." in: The journal of maternal-fetal & neonatal medicine, Vol. 29, Issue 19, pp. 3193-7, 2016
Forte, Torricelli, Boanini, Gazzano, Rubini, Fini, Bigi: "Antioxidant and bone repair properties of quercetin-functionalized hydroxyapatite: An in vitro osteoblast-osteoclast-endothelial cell co-culture study." in: Acta biomaterialia, Vol. 32, pp. 298-308, 2016
Jiang, Wang, Tang, Yao, Zhou: "Regulation of Viral Infection-induced Airway Remodeling Cytokine Production by the TLR3-EGFR Signaling Pathway in Human Bronchial Epithelial Cells." in: COPD, pp. 1-6, 2016
Xing, Gu, Ma, Ye: "Biglycan up-regulated vascular endothelial growth factor (VEGF) expression and promoted angiogenesis in colon cancer." in: Tumour biology, Vol. 36, Issue 3, pp. 1773-80, 2015
Lin, Li, Lin, Yu, Tu, Hua, Lin, Xu, Han, Chen: "Silencing of Livin inhibits tumorigenesis and metastasis via VEGF and MMPs pathway in lung cancer." in: International journal of oncology, Vol. 47, Issue 2, pp. 657-67, 2015
Xiao, Wu, Yin, Han, Ding, Qiao, Lu, Deng, Bo, Gong: "Wogonin Inhibits Tumor-derived Regulatory Molecules by Suppressing STAT3 Signaling to Promote Tumor Immunity." in: Journal of immunotherapy (Hagerstown, Md. : 1997), Vol. 38, Issue 5, pp. 167-84, 2015
Salamanna, Pagani, Maglio, Borsari, Giavaresi, Martelli, Buontempo, Fini: "Estrogen-deficient osteoporosis enhances the recruitment and activity of osteoclasts by breast cancer cells." in: Histology and histopathology, Vol. 31, Issue 1, pp. 83-93, 2015
Piotrowski, Kiszałkiewicz, Górski, Antczak, Górski, Pastuszak-Lewandoska, Migdalska-Sęk, Domańska-Senderowska, Nawrot, Czarnecka, Kurmanowska, Brzeziańska-Lasota: "Immunoexpression of TGF-β/Smad and VEGF-A proteins in serum and BAL fluid of sarcoidosis patients." in: BMC immunology, Vol. 16, pp. 58, 2015
Song, Han, Cai, Tang, Yang, Ao, Zhou: "The effects of self-assembling peptide RADA16 hydrogel on malignant phenotype of human hepatocellular carcinoma cell." in: International journal of clinical and experimental medicine, Vol. 8, Issue 9, pp. 14906-15, 2015
Xu, Zhou, Wang, Zhao, Li, Zhou, Su, Xu, Xia, Qian, Tu, Xiao, Chen, Chen, Wang: "Role of microRNA-27a in down-regulation of angiogenic factor AGGF1 under hypoxia associated with high-grade bladder urothelial carcinoma." in: Biochimica et biophysica acta, Vol. 1842, Issue 5, pp. 712-25, 2014
Li, Wang, Zhang, Zhao, Huang, Wu, Li, Li, Liu, Cao, Dai, Fang, Shang, Cao, Zhao, Chen: "Elevated PLGF contributes to small-cell lung cancer brain metastasis." in: Oncogene, Vol. 32, Issue 24, pp. 2952-62, 2013
Patschan, Patschan, Henze, Blaschke, Wessels, Müller: "Impairment and Differential Expression of PR3 and MPO on Peripheral Myelomonocytic Cells with Endothelial Properties in Granulomatosis with Polyangiitis." in: International journal of nephrology, Vol. 2012, pp. 715049, 2012
He, Xia, Wang, Wei, Luo, Li: "Multiple release of polyplexes of plasmids VEGF and bFGF from electrospun fibrous scaffolds towards regeneration of mature blood vessels." in: Acta biomaterialia, Vol. 8, Issue 7, pp. 2659-69, 2012
Hotowy, Sawosz, Pineda, Sawosz, Grodzik, Chwalibog: "Silver nanoparticles administered to chicken affect VEGFA and FGF2 gene expression in breast muscle and heart." in: Nanoscale research letters, Vol. 7, Issue 1, pp. 418, 2012
Larráyoz, de Luis, Rúa, Velilla, Cabello, Martínez: "Molecular effects of doxycycline treatment on pterygium as revealed by massive transcriptome sequencing." in: PLoS ONE, Vol. 7, Issue 6, pp. e39359, 2012
Nourshahi, Hedayati, Ranjbar: "The correlation between resting serum leptin and serum angiogenic indices at rest and after submaximal exercise." in: Regulatory peptides, Vol. 173, Issue 1-3, pp. 6-12, 2011
Goldberg, Prentice, Davies, Murgatroyd: "Overnight and basal metabolic rates in men and women." in: European journal of clinical nutrition, Vol. 42, Issue 2, pp. 137-44, 1988
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