Cusabio
ELISA Kit
Human Vascular Endothelial cell Growth Factor , VEGF ELISA Kit
This gene is a member of the PDGF/VEGF growth factor family and encodes a protein that is often found as a disulfide linked homodimer. This protein is a glycosylated mitogen that specifically acts on endothelial cells and has various effects, including mediating increased vascular permeability, inducing angiogenesis, vasculogenesis and endothelial cell growth, promoting cell migration, and inhibiting apoptosis. Elevated levels of this protein is linked to POEMS syndrome, also known as Crow-Fukase syndrome. Mutations in this gene have been associated with proliferative and nonproliferative diabetic retinopathy. Alternatively spliced transcript variants, encoding either freely secreted or cell-associated isoforms, have been characterized. There is also evidence for the use of non-AUG (CUG) translation initiation sites upstream of, and in-frame with the first AUG, leading to additional isoforms. [provided by RefSeq]
Product Name:

Human Vascular Endothelial cell Growth Factor , VEGF ELISA Kit

Product Type:
ELISA Kit
Code:
CSB-E11718h
Alias:
RP1-261G23.1, MGC70609, MVCD1, VEGF, VEGF-A, VPF, vascular endothelial growth factor isoform VEGF165|vascular permeability factor
Size:
96T
Species:
Human
other species: Bovine Chicken Dog Guinea pig Horse Mouse Pig Rat Sheep Hamster
Target Name:
vascular endothelial growth factor A
Abbreviation:
VEGFA
Protein Biological Process 1:
Angiogenesis
Protein Biological Process 3:
Angiogenesis
Detect Range:
31.25 pg/ml-2000 pg/ml
Sensitivity:
25.297pg/ml
Assay Time:
1-5h
Sample Volume:
50-100ul
Detection wavelengt:
450 nm
Download:

Protocol may be improved. Please feel free to contact CUSABIO product specialist to obtain the latest version.

Specificty: This assay has high sensitivity and excellent specificity for detection of Human VEGFA. No significant cross-reactivity or interference between Human VEGFA and analogues was observed.
Precision: Intra-assay Precision (Precision within an assay): CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10%
Three samples of known concentration were tested in twenty assays to assess.
Sample collection and storage: Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 4°C before centrifugation for 15 minutes at 1000 ×g. Remove serum and assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles.
Plasma: Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 ×g at 2-8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles.
Assay procedure: Bring all reagents and samples to room temperature before use. Centrifuge the sample again after thawing before the assay. It is recommended that all samples and standards be assayed in duplicate.
1. Prepare all reagents, working standards, and samples as directed in the previous sections.
2. Refer to the Assay Layout Sheet to determine the number of wells to be used and put any remaining wells and the desiccant back into the pouch and seal the ziploc, store unused wells at 4°C.
3. Add 100μl of standard and sample per well. Cover with the adhesive strip provided. Incubate for 2 hours at 37°C. A plate layout is provided to record standards and samples assayed.
4. Remove the liquid of each well, don't wash.
5. Add 100μl of Biotin-antibody (1x) to each well. Cover with a new adhesive strip. Incubate for 1 hour at 37°C. (Biotin-antibody (1x) may appear cloudy. Warm up to room temperature and mix gently until solution appears uniform.)
6. Aspirate each well and wash, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (200μl) using a squirt bottle, multi-channel pipette, manifold dispenser, or autowasher, and let it stand for 2 minutes, complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining wash Buffer by aspirating ordecanting. Invert the plate and blot it against clean paper towels.
7. Add 100μl of HRP-avidin (1x) to each well. Cover the microtiter plate with a new adhesive strip. Incubate for 1 hour at 37°C.
8. Repeat the aspiration/wash process for five times as in step 6.
9. Add 90μl of TMB Substrate to each well. Incubate for 15-30 minutes at 37°C. Protect from light.
10. Add 50μl of Stop Solution to each well, gently tap the plate to ensure thorough mixing.
11. Determine the optical density of each well within 5 minutes, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. Subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
Calculation of results: Using the professional soft "Curve Expert 1.3" to make a standard curve is recommended, which can be downloaded from our web.
Average the duplicate readings for each standard and sample and subtract the average zero standard optical density.
Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the x-axis against the concentration on the y-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the VEGFA concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data.
If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.

Literature1Literature2Literature3Literature4Literature5Literature6Literature7Literature8

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AuthorTang YT et al
PMID23137522
Linkhttp://www.ncbi.nlm.nih.gov/pubmed/?term=23137522
Type of publicationProduct-specific
Year2012
Impact Factor2.923
JournalLeuk Res
Other information2013 Apr;37(4):359-66. doi: 10.1016/j.leukres.2012.10.008. Epub 2012 Nov 6.
TitlePreparation and characterization of decellularized tendon slices for tendon tissue engineering
Linkhttp://onlinelibrary.wiley.com/doi/10.1002/jbm.a.34083/full
Type of publicationProduct-specific
Impact Factor2.625
Journaljournal of biomedical materials research part a
TitlePropranolol given orally for proliferating infantile haemangiomas: analysis of efficacy and serological changes in vascular endothelial growth factor and endothelial
AuthorYuan WL et al
PMID23291092
Linkhttp://www.ncbi.nlm.nih.gov/pubmed/?term=23291092
Type of publicationProduct-specific
Year2013
Impact Factor1.95
JournalBr J Oral Maxillofac Surg
Other information2013 Jan 3. pii: S0266-4356(12)00623-7. doi: 10.1016/j.bjoms.2012.12.003. [Epub ahead of print]
TitleThe biological effects and mechanisms of calcitonin gene-related peptide on human endothelial cell
AuthorTuo Y et al
PMID23461295
Linkhttp://www.ncbi.nlm.nih.gov/pubmed/?term=23461295
Type of publicationProduct-specific
Year2013
Impact Factor1.588
JournalJ Recept Signal Transduct Res.
Other information2013 Apr;33(2):114-23. doi: 10.3109/10799893.2013.770528. Epub 2013 Mar 6.
TitleThe Role of Insulin-Like Growth Factor I and Hypoxia Inducible Factor 1α in Vascular Endothelial Growth Factor Expression in Type 2 Diabetes
AuthorJiang F et al
PMID23462604
Linkhttp://www.ncbi.nlm.nih.gov/pubmed/?term=23462604
Type of publicationProduct-specific
Year2013
Impact Factor0.956
JournalAnn Clin Lab Sci.
Other information2013 Winter;43(1):37-44.
TitleEpigenetic silencing of miR-126 contributes to tumor invasion and angiogenesis in colorectal cancer
AuthorZhang Y et al
PMID23900443
Linkhttp://www.ncbi.nlm.nih.gov/pubmed/23900443
Type of publicationProduct-specific
Year2013
Impact Factor2.297
JournalOncol Rep
Other information2013 Jul 23. doi: 10.3892/or.2013.2633. [Epub ahead of print]
TitleMicroRNA
AuthorYang X et al
PMID24259426
Linkhttp://www.ncbi.nlm.nih.gov/pubmed/24259426
Type of publicationProduct-specific
Year2013
Impact Factor12.003
JournalHepatology
Other information2013 Nov 21. doi: 10.1002/hep.26941. [Epub ahead of print]
TitleThe Association of Serum Vascular Endothelial Growth Factor and Ferritin in Diabetic Microvascular Disease
AuthorGuo L et al
PMID24279470
Linkhttp://www.ncbi.nlm.nih.gov/pubmed/24279470
Type of publicationProduct-specific
Year2013
Impact Factor2.205
JournalDiabetes Technol Ther
Other information2013 Nov 26. [Epub ahead of print]