Human VEGF PicoKine™ ELISA Kit

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SKU EK0539
Size 96wells/kit, with removable strips.
Reactivity Human
Sample Type cell culture supernates, serum and plasma(heparin, EDTA, citrate).
Sensitivity <1pg/ml
Assay Range 31.2pg/ml-2000pg/ml(humanserum,plasma)15.6pg/ml-1000pg/ml(cellculturesupernates)

Overview

Product Name Human VEGF PicoKine™ ELISA Kit
SKU/Catalog Number EK0539
Storage & Handling Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.)
Size 96wells/kit, with removable strips.
Description Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human VEGF. 96wells/kit, with removable strips.
Cite This Product Human VEGF PicoKine™ ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0539)
Sample Type cell culture supernates, serum and plasma(heparin, EDTA, citrate).. Anticoagulant(s): heparin, EDTA or citrate
*The recommended anticoagulants are proven to not block the antibody binding sites on the target antigen. Please do not collect blood sample with other anticoagulants that are not specified above or contact us to check for feasibility.
Sensitivity <1pg/ml
Assay Range 31.2pg/ml-2000pg/ml(humanserum,plasma)15.6pg/ml-1000pg/ml(cellculturesupernates)
Immunogen Expression system for standard: sf21; Immunogen sequence: A27-R191
Reactivity Human
Cross Reactivity There is no detectable cross-reactivity with other relevant proteins.
Antibody Clonalities Capture Antibody | Detection Antibody:
monoclonal antibody from mouse|polyclonal antibody from goat

Assay Details

Kit Components

Catalog numberDescriptionQuantity
EK0539-CAP96-well plate precoated with anti-Human VEGFA antibody1
EK0539-STlyophilized recombinant Human VEGFA standard10ng/tube
EK0539-DAbiotinylated anti-Human VEGFA antibody130ul
AR1103Avidin-Biotin-Peroxidase Complex(ABC)130ul
AR1106-1sample diluent buffer30ml
AR1106-2antibody diluent buffer12ml
AR1106-3ABC diluent buffer12ml
AR1104TMB color developing agent10ml
AR1105TMB stop solution10ml
PLA-SEAAdhesive cover4

*Why there is no wash buffer? Our Avidin-Biotin-Peroxidase Diluent contains the detergent (TWEEN) normally present in other companies' ELISA kits. This saves you the step of having to wash with the special wash buffer and achieve similar or better signal to noise ratio. The wash can use regular wash buffers (PBS, TBS etc.) commonly found in labs.

Materials Required But Not Provided

  • Microplate reader in standard size.
  • Automated plate washer.
  • Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
  • Clean tubes and Eppendorf tubes.
  • Washing buffer (neutral PBS or TBS).
  • Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g NaCl; 450μl of purified acetic acid or 700μl of concentrated hydrochloric acid to 1000ml H2
  • Preparation of 0.01 M PBS: Add 8.5g sodium chloride, 1.4g Na O and adjust pH to 7.2-7.

Typical Data Obtained from Human VEGF PicoKine™ ELISA Kit

(TMB reaction incubate at 37°C for 15-25min)

Concentration(pg/ml)031.262.512525050010002000
O.D.0.0200.1130.1890.3450.5890.9901.6062.313

Intra/Inter Assay Precision

Intra-Assay PrecisionInter-Assay Precision
Sample123123
n161616242424
Mean(pg/ml)7227094676254955
Standard deviation4.5321.3359.599.6224.3874.49
CV(%)9.6%7.9%9.6%7.8%9.6%7.8%

Reproducibility

Three samples with differing target protein concentrations were assayed using four different lots to measure the CV% lot to lot variance.

Reproducibility

To assay reproducibility, three samples with differing target protein concentrations were assayed using four different lots.

LotsLot1 (pg/ml)Lot2 (pg/ml)Lot3 (pg/ml)Lot4 (pg/ml)Mean (pg/ml)Standard DeviationCV (%)
Sample 172808271764.816.3%
Sample 227026725224525810.354%
Sample 39468971026103797657.795.9%
*number of samples for each test n=16.

*The typical data is obtained from Boster's internal QC result and for reference only. It may differ from the lab test results of the end users. It is more important that the user's lab test results reflect the same linearity demonstrated in the typical data than achieving exactly the same O.D. values.

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id P15692
Gene Name VEGFA
Protein Name Vascular endothelial growth factor A
Tissue Specificity Isoform VEGF189, isoform VEGF165 and isoform VEGF121 are widely expressed. Isoform VEGF206 and isoform VEGF145 are not widely expressed. A higher level expression seen in pituitary tumors as compared to the pituitary gland. .
Alternative Names Vascular endothelial growth factor A;VEGF-A;Vascular permeability factor;VPF;VEGFA;VEGF;
Subcellular Localization Secreted . VEGF121 is acidic and freely secreted. VEGF165 is more basic, has heparin-binding properties and, although a signicant proportion remains cell-associated, most is freely secreted. VEGF189 is very basic, it is cell-associated after secretion and is bound avidly by heparin and the extracellular matrix, although it may be released as a soluble form by heparin, heparinase or plasmin.
Molecular Weight 27042 MW

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Protein Function Growth factor active in angiogenesis, vasculogenesis and endothelial cell growth. Induces endothelial cell proliferation, promotes cell migration, inhibits apoptosis and induces permeabilization of blood vessels. Binds to the FLT1/VEGFR1 and KDR/VEGFR2 receptors, heparan sulfate and heparin. NRP1/Neuropilin-1 binds isoforms VEGF-165 and VEGF-145. Isoform VEGF165B binds to KDR but does not activate downstream signaling pathways, does not activate angiogenesis and inhibits tumor growth. .
Research Areas Angiogenesis, Cancer, Cancer Metabolism, Cardiovascular, Growth Factors, Growth Factors/Hormones, Metabolism, Metabolism Processes, Pathways And Processes, Response To Hypoxia, Signal Transduction

*You can search these to find other products in these research areas.
Background Vascular permeability factor/vascular endothelial growth factor(VPF/VEGF), a potent cytokine expressed by most malignant tumors, has critical roles in vasculogenesis and both physiological and pathological angiogenesis.VEGF produced by tumor cells potently stimulates endothelial cell proliferation and angiogenesis and plays a key role in the pathophysiology of several neoplasias. VEGF may also play a pivotal role in mediating the development and progression of diabetic retinopathy.VEGF, a major regulator of angiogenesis, binds to two receptor tyrosine kinases, KDR/Flk-1 and Flt-1. The VEGF gene is mapped by fluorescence in situ hybridization to chromosome 6p12.

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Publications

Histone demethylase KDM4D promotes gastrointestinal stromal tumor progression through HIF1?/VEGFA signalling
PubMed ID30060750
The expression and underlying angiogenesis effect of DPC4 and VEGF on the progression of cervical carcinoma
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Growth Factor Variation in Two Types of Autologous Platelet Biomaterials: PRP Versus PRF
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DHX32 Promotes Angiogenesis in Colorectal Cancer Through Augmenting ?-catenin Signaling to Induce Expression of VEGFA
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High fibrosis indices in cerebrospinal fluid of patients with shunt-dependent post-traumatic chronic hydrocephalus
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5-Aminolaevulinic Acid-Based Photodynamic Therapy Restrains Pathological Hyperplasia of Fibroblasts
PubMed ID28052053
Elevated expression levels of serum insulin-like growth factor-1, tumor necrosis factor-? and vascular endothelial growth factor 165 might exacerbate type 2 diabetic nephropathy
PubMed ID27218216
Genetic Risk Factors for Psoriasis in Turkish Population: -1540 C/A, -1512 Ins18, and +405 C/G Polymorphisms within the Vascular Endothelial Growth Factor Gene
PubMed ID26848216
Wogonin inhibits multiple myeloma-stimulated angiogenesis via c-Myc/VHL/HIF-1? signaling axis
PubMed ID26735336
Human umbilical cord mesenchymal stem cells promote peripheral nerve repair?via?paracrine mechanisms
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Serum level of vascular endothelial growth factor decreased in chronic ketamine abusers
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CXCR7 stimulates MAPK signaling to regulate hepatocellular carcinoma progression
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Effects of laser photocoagulation on serum angiopoietin-1, angiopoietin-2, angiopoietin-1/angiopoietin-2 ratio, and soluble angiopoietin receptor Tie-2 levels in type 2 diabetic patients with proliferative diabetic retinopathy
PubMed ID25161936
Roles of PI3K/Akt and c-Jun Signaling Pathways in Human Papillomavirus Type 16 Oncoprotein-Induced HIF-1?, VEGF, and IL-8 Expression and?In Vitro?Angiogenesis in Non-Small Cell Lung Cancer Cells
PubMed ID25058399
Evaluating the Potential Bioactivity of a Novel Compound ER1626
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MicroRNA-26a Inhibits Angiogenesis by Down-Regulating VEGFA through the PIK3C2?/Akt/HIF-1? Pathway in Hepatocellular Carcinoma
PubMed ID24194905
Radioactive 125I Seed Inhibits the Cell Growth, Migration, and Invasion of Nasopharyngeal Carcinoma by Triggering DNA Damage and Inactivating VEGF-A/ERK Signaling
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Overexpression of HTRA1 Leads to Down-Regulation of Fibronectin and Functional Changes in RF/6A Cells and HUVECs
PubMed ID23056244
Effect of essential fatty acids on glucose-induced cytotoxicity to retinal vascular endothelial cells
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Development of small interfering RNA delivery system using PEI-PEG-APRPG polymer for antiangiogenic vascular endothelial growth factor tumor-targeted therapy
PubMed ID21904456
Inhibitory effect of vascular endothelial growth factors-targeted small interfering RNA on proliferation of gastric cancer cells.
PubMed ID17465445
Development of small interfering RNA delivery system using PEI-PEG-APRPG polymer for antiangiogenic vascular endothelial growth factor tumor-targeted therapy
PubMed ID21904456
Inhibitory effect of vascular endothelial growth factors-targeted small interfering RNA on proliferation of gastric cancer cells.
PubMed ID17465445
A Y, Li Y, Zhao S. Oncol Lett. 2018 Feb;15(2):2534-2540. doi: 10.3892/ol.2017.7580. Epub 2017 Dec 11. The expression and underlying angiogenesis effect of DPC4 and VEGF on the progression of cervical carcinoma
PubMed ID29434970
Ma X, Li Z, Li T, Zhu L, Li Z, Tian N. Am J Transl Res. 2017 Nov 15;9(11):5012-5021. eCollection 2017. Long non-coding RNA HOTAIR enhances angiogenesis by induction of VEGFA expression in glioma cells and transmission to endothelial cells via glioma cell derived-extracellular vesicles
PubMed ID29218099
Wang X, Cao P, Liu J, Du P, Wang Z, Chen W, Liu C, Wu Y. Med Sci Monit. 2017 Jan 4;23:46-56. 5-Aminolaevulinic Acid-Based Photodynamic Therapy Restrains Pathological Hyperplasia of Fibroblasts
PubMed ID28052053
Xiaochuan Wang, Ping Cao, Jian Liu, Peng Du, Zhiqiong Wang, Wei Chen, Chang Liu, Yifei Wu (Department of Dermatology, The First People’s Hospital of Yunnan Province, Kunming, Yunnan, China (mainland)) Med Sci Monit 2017; 23:46-56 DOI: 10.12659/MSM.898221 Published: 2017-01-04 5-Aminolaevulinic Acid-Based Photodynamic Therapy Restrains Pathological Hyperplasia of Fibroblasts
PubMed ID-
Xiaochuan Wang, Ping Cao, Jian Liu, Peng Du, Zhiqiong Wang, Wei Chen, Chang Liu, Yifei Wu (Department of Dermatology, The First People’s Hospital of Yunnan Province, Kunming, Yunnan, China (mainland)) Med Sci Monit 2017; 23:46-56 DOI: 10.12659/MSM.898221 Published: 2017-01-04 5-Aminolaevulinic Acid-Based Photodynamic Therapy Restrains Pathological Hyperplasia of Fibroblasts
PubMed ID-
Wu H, Deng J, Yu S, Wang X, Chen Y. J Huazhong Univ Sci Technolog Med Sci. 2010 Feb;30(1):108-12. Doi: 10.1007/S11596-010-0120-6. Epub 2010 Feb 14. The Inhibitory Effects Of Rh-Endostatin (Yh-16) In Combination With Radiotherapy On Lung Adenocarci...
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Chuangui C, Peng T, Zhentao Y. Pathol Oncol Res. 2012 Oct;18(4):1021-7. Epub 2012 Apr 29. The Expression Of High Mobility Group Box 1 Is Associated With Lymph Node Metastasis And Poor Prognosis In Esophageal Squamous Cell Carcinoma.
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Xu Yf, Ge Fj, Han B, Yang Xq, Su H, Zhao Ac, Zhao Mh, Yang Yb, Yang J. World J Gastroenterol. 2015 Mar 21;21(11):3256-65. Doi: 10.3748/Wjg.V21.I11.3256. High-Mobility Group Box 1 Expression And Lymph Node Metastasis In Intrahepatic Cholangiocarcin...
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Yang W, Zhang Y, Yu J, Li S. Eur J Surg Oncol. 2010 May;36(5):501-6. Doi: 10.1016/J.Ejso.2010.01.007. Epub 2010 Feb 23. The Low Expression Of Cd80 Correlated With The Vascular Endothelial Growth Factor In Esophageal Cancer Tissue.
PubMed ID20181455
Zhou L, Zhang K, Li J, Cui X, Wang A, Huang S, Zheng S, Lu Y, Chen W. Reprod Toxicol. 2013 Jun;37:62-9. Doi: 10.1016/J.Reprotox.2013.02.001. Epub 2013 Feb 11. Inhibition Of Vascular Endothelial Growth Factor-Mediated Angiogenesis Involved In Repro...
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Chen J, Wei D, Zhuang H, Qiao Y, Tang B, Zhang X, Wei J, Fang S, Chen G, Du P, Huang X, Jiang W, Hu Q, Hua Zc. Mol Cell Proteomics. 2011 Jun;10(6):M111.009399. Doi: 10.1074/Mcp.M111.009399. Epub 2011 Apr 7. Proteomic Screening Of Anaerobically Reg...
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Fu Z, Chen D, Cheng H, Wang F. Med Sci Monit. 2015 Jan 27;21:318-25. Doi: 10.12659/Msm.893265. Hypoxia-Inducible Factor-1?? Protects Cervical Carcinoma Cells From Apoptosis Induced By Radiation Via Modulation Of Vascular Endothelial Growth Factor A...
PubMed ID25623525
Liu Y, Lv L. J Huazhong Univ Sci Technolog Med Sci. 2004;24(5):470-2. Mechanism Of Elevated Vascular Endothelial Growth Factor Levels In Peritoneal Fluids From Patients With Endometriosis.
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Li Xy, Lin Yc, Huang Wl, Hong Cq, Chen Jy, You Yj, Li Wb. Med Oncol. 2012 Jun;29(2):714-20. Doi: 10.1007/S12032-011-9904-1. Epub 2011 Mar 24. Zoledronic Acid Inhibits Proliferation And Impairs Migration And Invasion Through Downregulating Vegf And...
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Ye X, Ren H, Zhang M, Sun Z, Jiang Ac, Xu G. Invest Ophthalmol Vis Sci. 2012 Jun 8;53(7):3481-9. Doi: 10.1167/Iovs.11-9076. Erk1/2 Signaling Pathway In The Release Of Vegf From M??ller Cells In Diabetes.
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Shen J, Shen S, Das Un, Xu G. Lipids Health Dis. 2012 Jul 10;11:90. Doi: 10.1186/1476-511X-11-90. Effect Of Essential Fatty Acids On Glucose-Induced Cytotoxicity To Retinal Vascular Endothelial Cells.
PubMed ID22781401
Liu D, Cao G, Cen Y, Liu T, Peng W, Sun J, Li X, Zhou H. Int Immunopharmacol. 2013 Oct;17(2):237-44. Doi: 10.1016/J.Intimp.2013.06.002. Epub 2013 Jun 19. The Radiosensitizing Effect Of Cpg Odn107 On Human Glioma Cells Is Tightly Related To Its Ant...
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Chai Zt, Kong J, Zhu Xd, Zhang Yy, Lu L, Zhou Jm, Wang Lr, Zhang Kz, Zhang Qb, Ao Jy, Wang M, Wu Wz, Wang L, Tang Zy, Sun Hc. Plos One. 2013 Oct 23;8(10):E77957. Doi: 10.1371/Journal.Pone.0077957. Ecollection 2013. Microrna-26A Inhibits Angiogenes...
PubMed ID24194905
Li Xq, Ouyang Zg, Zhang Sh, Liu H, Shang Y, Li Y, Zhen Ys. Cancer Biol Ther. 2014 Apr;15(4):398-408. Doi: 10.4161/Cbt.27626. Epub 2014 Jan 14. Synergistic Inhibition Of Angiogenesis And Glioma Cell-Induced Angiogenesis By The Combination Of Temozo...
PubMed ID24424202
Wang L, Zeng Y, Wang T, Liu H, Xiao H, Xiang H. Plos One. 2014 Jan 27;9(1):E86509. Doi: 10.1371/Journal.Pone.0086509. Ecollection 2014. Evaluating The Potential Bioactivity Of A Novel Compound Er1626.
PubMed ID24475135
Zhang H, Chen Y, Fan B, Wang W, Zhu W. Tumour Biol. 2015 May;36(5):3871-80. Doi: 10.1007/S13277-014-3029-Z. Epub 2015 Jan 11. Overexpression Of Vegf183 Promotes Murine Breast Cancer Cell Proliferation In Vitro And Induces Dilated Intratumoral Micr...
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Das Kk, Jargar Jg, Saha S, Yendigeri Sm, Singh Sb. Indian J Pharmacol. 2015 May-Jun;47(3):285-91. Doi: 10.4103/0253-7613.157126. ??-Tocopherol Supplementation Prevents Lead Acetate And Hypoxia-Induced Hepatic Dysfunction.
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Fan N, Zhang M, Xu K, Ke X, Ding Y, Wang D, Liu Y, Ning Y, Deng X, He H. Drug Alcohol Depend. 2015 Jul 1;152:57-61. Doi: 10.1016/J.Drugalcdep.2015.04.022. Epub 2015 May 8. Serum Level Of Vascular Endothelial Growth Factor Decreased In Chronic Keta...
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Xu Wh, Ge Yl, Li Q, Zhang X, Duan Jh. World J Gastroenterol. 2007 Apr 14;13(14):2044-7. Inhibitory Effect Of Vascular Endothelial Growth Factors-Targeted Small Interfering Rna On Proliferation Of Gastric Cancer Cells.
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En-Lin S, Sheng-Guo C, Hua-Qiao W. Gynecol Oncol. 2010 Jun;117(3):417-22. Doi: 10.1016/J.Ygyno.2009.12.016. Epub 2010 Apr 7. The Expression Of Efemp1 In Cervical Carcinoma And Its Relationship With Prognosis.
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He W, Guo Z, Wen Y, Wang Q, Xie B, Zhu S, Wang Q. J Biomater Sci Polym Ed. 2012;23(1-4):315-31. Doi: 10.1163/092050610X550359. Epub 2011 Jan 18. Alginate-Graft-Pei As A Gene Delivery Vector With High Efficiency And Low Cytotoxicity.
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Song El, Hou Yp, Yu Sp, Chen Sg, Huang Jt, Luo T, Kong Lp, Xu J, Wang Hq. Gynecol Oncol. 2011 Apr;121(1):174-80. Doi: 10.1016/J.Ygyno.2010.11.004. Epub 2010 Dec 15. Efemp1 Expression Promotes Angiogenesis And Accelerates The Growth Of Cervical Can...
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Zhang E, Feng X, Liu F, Zhang P, Liang J, Tang X. Plos One. 2014 Jul 24;9(7):E103440. Doi: 10.1371/Journal.Pone.0103440. Ecollection 2014. Roles Of Pi3K/Akt And C-Jun Signaling Pathways In Human Papillomavirus Type 16 Oncoprotein-Induced Hif-1??, V...
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Customer Q&As

Q: The volumes of my samples are small, would it be possible to use 50µl/sample?
A: There are two solutions for insufficient sample volume.
1. Diluent the samples with the provided sample diluent buffer into 100ul.
2. Add 50ul of standard solution, when 50ul of sample will be added into a well.
Q: Will this kit work on tissue samples?
A: For tissue homogenates, endogenous biotin should be considered. Endogenous biotin in tissue homogenates might introduce false positive signal. Please run the test as described below to confirm if there is Endogenous biotin in the tissue lysate samples using the ELISA kit.
• Add tissue homogenates into the wells and then add ABC and TMB without adding any biotinylated detection antibody to see if any signal will be observed.
• If no signal is produced, then you can work on the tissue sample by using the kit.
Q: Is it possible to dilute 10X TBS stock that is 250mM Tris Base/Tris-Hcl, 27 mM KCl, and 1.37M NaCl to be used as the wash buffer?
A: You can dilute the 10X TBS stock to 1XTBS and test the pH value of the 1xTBS.
The 1XTBS can be used if the pH value falls in the range of 7.2-7.6.
Q: Would your kits work normally with samples from Wistar Rat?
A: Our kits would work normally with samples from Wistar Rat.
Q: How can I store the dissolved wash buffer for a longer period of time?
A: You can prepare a concentrated wash buffer (25X) which can be stored at 4°C for 1-3 months.
Q: Does the TMB color developing agent contain H2O2?
A: Yes, the TMB color developing agent contain H2O2.
Q: Does this ELISA kit contain any product produced in humans or primates?
Keywords: component, ingredient
A: None of the components in our ELISA kits are produced in humans or primates.
Q: What is the volume of the recombinant protein control? What is its shelf life?
Keywords: expiration, storage, temperature, size
A: The volume of the control is around 100pg. If unopened, the shelf life of the control is the same as the whole kit - "Store at 4˚C for 6 months, at -20˚C for 12 months." The reconstituted control can be stored at -20˚C for 2 days.
Q: Will it be okay to run the experiment if I accidentally used the wrong buffer (e.g. sample diluent buffer) for antibody dilution instead of the antibody diluent buffer? Keywords: dilution, replace, substitute, sample buffer
A: Using the sample diluent buffer for antibody dilution instead of the antibody diluent buffer will negatively affect the test result to some extent. The values of both standard and sample might be lower than the normal values.
Q: The protocol says to use neutral PBS and provides a recipe. Could I use neutral PBS made with KCl or KH2PO4 (common constituents for most PBS recipes) instead? Keywords: protocol, alternative buffer, applications, reagent recipe
A: Yes, it is ok to use common PBS recipes. We have tried many types of buffers with common constituents, and no significant difference was observed whatsoever.
Q: Why are my O.D. values different than your values on the datasheet? Keyword: troubleshooting, reference
A: We detected the kit in our lab and got our values on the datasheet before delivery, but protein activity will decrease as time goes on, so you may get lower O.D. values than ours. However, it should still be in a reasonable range and the standards can be used to calculate sample values. Good linearity of curve is more important than the actual numerical value.
Q: On the ELISA kit datasheet it says "HRP substrate TMB was used to visualize HRP enzymatic reaction." What is HRP and which part of the kit contains HRP? Keyword: kit components, reagents, protocol
A: It means Avidin-Biotin-Peroxidase Complex(AR1103) and you could find it in Kit Components.
Q: Do you offer any ELISA kits that can work with whole blood samples instead of plasma or urine? Keyword: applications
A: We test serum and plasma routinely, and there is very little difference between serum, plasma and whole blood. The whole blood also contains proteins we need to test. The kit can be used to test whole blood in theory.
Q: Does silicic acid (formula SiOH4) affect the results of the ELISA assay? Keyword: protocol, reagents
A: The acid may affect the binding of antigen to antibody, it is not recommended to use. That being said, it is unlikely to affect the reaction if the solution remains an overall neutral environment.
Q: Is there lot-to-lot variation of the ELISA kits? What are Boster's general services when I have questions about your kits? Keyword: technical support, help, customer service
A: The NIBSC/WHO 1st International Standard is evaluated, we always test our kits before delivery, and customers can find the test result on the protocol. If the customer needs technical support from us to analyze their data, please contact us and we ask that you provide us the following information: the lot# and production date, and when did the customer detect the kit.
Q: The results of my standards do not look correct, what could be the problem? Keyword: troubleshoot, protocol
A: Double check the protocol for plate washing. It should be examined on three aspects: 1) Did you make the washing buffer correctly? 2) How long did you let the buffer stay in wells? 3) What was the volume of the washing buffer added to each well? In addition, pay attention when adding sample to avoid contamination. And we suggest testing the standards again. Values of standards at low concentration are more affected than that at higher concentration, so customer can still get expected values at high concentration even if there is an error. If problems persist, please contact technical support with the catalog and lot number of your product.
Q: What is the well depth of the 96 well plates for the ELISA kits? Keyword: well capacity, product size
A: The well depth is 300ul, and the max capacity is 350ul. The height of the well is 1.1 cm, and the internal size is 1 cm.
Q: The kit does not include wash solution, what should I do? Keyword: wash buffer
A: Our Elisa kits do not come with wash solution, but we have included information about how to make washing buffer on the datasheet. Please refer to the "Material Required But Not Provided" section. We also offer washing buffer for sale separately (Phosphate Buffered Saline Powder SKU: AR0030).
Q: For your ELISA kits, are the capture and detection antibodies polyclonals or monoclonals? Keyword: antibody clonality
A: This information can be found for each kit under the "Properties" section, and you can find the immunogen sequence information in the "Overview" section.
Q: Do your ELISA kits come with sealants or plate covers? Keyword: storage, sequential use
A: The kit can be used within a month sequentially if it's opened and stored at 4 degree. There is no need to use sealants, the plate can be packaged with aluminum foil bag, and for other reagents keep bottle tightly closed.
Q: Are the mouse, rat, and human VEGF kits truly specific for each species or do they cross react? Which isoforms of VEGF do the human, mouse and rat kit detect? Keyword: specificity
A: Our VEGF Elisa kits are truly specific for each species, and they detect 165 amino acid form.
Q: What is the concentration of Sodium Azide in your Elisa kits? Keyword: preservative
A: Our Elisa kits contain 0.02% Sodium Azide. All of the components except TMB colour developing reagent and stop solution contains 0.02% Sodium azide as the preservative.
Q: Are there positive and negative controls available for my ELISA kit? Keyword: positive control, negative control
A: We can provide a recombinant protein as a control. It costs $50 per control and takes 2 weeks to manufacture. We cannot provide a positive or negative control in serum.
Q: Why do I get positive results for my knockout (KO) model when used as a control?
A: The knockout (KO) model may contain truncated forms of the target protein which can be detected by the ELISA assay.
Q: How long do I soak my plate in the wash buffer?
A: The plate should soak in 0.3mL PBS or TBS wash buffer for at least 1-2 minutes in an automated wash station.
Q: Can I use Tween in my wash buffer?
A: While it is not recommended to use Tween in your wash buffer, small amounts (<0.1% concentration) may decrease background due to insufficient blocking.
Q: What plate type do I use to set up the microplate reader?
A: Our plates are made with the Corning costar plates similar to the DNA-BIND 96 -well plates.
Q: Does this kit detect all forms of VEGF or just VEGFA
A: This assay detects VEGFA
Q: How come I saw no signal for running your kit with my VEGF plasma sample?
A: First of all, the amount of VEGF antigen in plasma is much lower than that in serum, and it is possible that the antigen is undetectable by the assay. You may use serum instead of plasma if applicable.
Q: Can you tell me how to troubleshoot?
A: To troubleshoot, you should try to repeat your experiment without diluting your plasma samples. If this approach still does not work, try to increase your sample incubation time to 2.5 hours (at room temperature). To minimize any potential sample losses, we recommend you pilot your experiment using standards and a small number of samples.
Q: What should I use for negative control?
A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the normal level of this protein in my sample of interest?
A: We have reviewed literature and have gathered this information for most of our ELISA kits. You can find this information on the product page or contact us if you cannot find it. However this information is only suggestive and cannot replace pilot studies in determining the optimal sample dilution ratio.
Q: Is the plate separable? Can I use only part of the kit and save the rest for later? How many samples can I run with one kit?
A: Yes the plate is separable. There are 12 strips of 8 wells each, all removable from the plate. The amount of samples you can run depend on a few factors. In the most common ELISA set up, you will use two strips for standards, and 10 strips for samples using duplicates, which let you run up to 40 samples per kit. Contact us if you have questions regarding other situations.
Q: Does the kit contain sample preparation reagents? How do I prepare my samples?
A: Since different sample types require different reagents, we do not include them in the ELISA kit. However we do have each reagent mentioned in the file below available at very reasonable prices. Be sure to check them out. For sample preparation protocols please download the file below: https://www.bosterbio.com/media/pdf/Boster_Sample_Preparation_Protocols.pdf
Q: Can this ELISA kit work on brain tissue homogenate, cell culture supernatant, saliva, urine, serum, whole blood or any other sample type?
A: In theory the ELISA kit will work for all sample types if the target protein is present at a level that falls within the linear range of the ELISA kit detection range. We guarantee the kit to work on the sample types that we have tested. If you want dilution ratio suggestions on these sample types please contact our technical support. For sample types that we have not tested for, we suggest you run pilot experiments to decide the optimal sample dilution.
Q: Can this ELISA kit react with the pro-form of the target protein? Can this ELISA kit react with an isoform of the protein?
A: In general, unless otherwise specified, the ELISA kit is pan-specific, meaning that it will react with all different forms of the target protein if they share the majority of the target protein's sequence. The capture and detection antibodies are reactive to the entire sequence of the standard protein. You can find the sequence information of the standard protein in the "immunogen" section. For more information about the specificity of the kit for your particular experiment, please contact our technical support.
Q: Can this ELISA kit react with human, mouse, rat or other species?
A: If the kit is reactive to another commonly used species (human, mouse, and/or rat), we would have listed it as a separate product. If you want to check cross-reactivity to a species that is not included in the 3 species listed above, please contact our technical support for more information. As a rule of thumb, if the sequences are 90%+ identical, there is a high chance of cross-reactivity for your species of interest.
Q: What are some alternative names that could be used to describe this product?
A: Some common names include but are not limited to human vegf a elisa kit, human vegfa elisa kit