Product Info Summary
SKU: | EK0539 |
---|---|
Size: | 96 wells/kit, with removable strips. |
Reactive Species: | Human |
Application: | ELISA |
Sample Types: | cell culture supernatants, serum and plasma (heparin, EDTA, citrate). |
Product info
Product Name
Human VEGF ELISA Kit PicoKine®
SKU/Catalog Number
EK0539
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human VEGF ELISA Kit PicoKine® (96 Tests). Quantitate Human VEGFA in cell culture supernatants, serum and plasma (heparin, EDTA, citrate). Sensitivity: 1pg/ml. The brand Picokine indicates this is a premium quality ELISA kit. Each Picokine kit delivers precise quantification, high sensitivity, and excellent reproducibility. Only our most reliable and effective kits qualify as Picokine, guaranteeing top-tier results for your assays.
Storage & Handling
Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)
Cite This Product
Human VEGF ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0539)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: NS0; Immunogen sequence: A27-R191
Sensitivity
<1 pg/ml
Assay Range
31.2 pg/ml - 2,000 pg/ml
Standard Dilution Instructions
See datasheet of EK0539 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK0539 is reactive to VEGFA in Human samples
Validated Sample Types
cell culture supernatants, serum and plasma (heparin, EDTA, citrate).
Application Guarantee
EK0539 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of VEGFA
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.
Kit Components
Catalog Number | Description | Quantity |
---|---|---|
EK0539-CAP | Anti-Human VEGFA Pre-coated 96-well strip microplate | 1 |
EK0539-ST | Human VEGFA Standard | 2 vials, 10 ng/tube |
EK0539-DA | Human VEGFA Biotinylated antibody (100x) | 100ul |
AR1103 | Avidin-Biotin-Peroxidase Complex (100x) | 100ul |
AR1106-1 | Sample Diluent | 30ml |
AR1106-2 | Antibody Diluent | 12ml |
AR1106-3 | Avidin-Biotin-Peroxidase Diluent | 12ml |
AR1104 | Color Developing Reagent (TMB) | 10ml |
AR1105 | Stop Solution | 10ml |
AR1106-5 | Wash Buffer (25x) | 20ml |
PLA-SEA | Adhesive plate sealers | 4 |
*The kit components are not available for individual purchase.
Materials Required But Not Included In Kit
- Microplate Reader capable of reading absorbance at 450nm.
- Incubator.
- Automated plate washer (optional).
- Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
- Multichannel pipettes are recommended for large amount of samples.
- Deionized or distilled water.
- 500ml graduated cylinders.
- Test tubes for dilution.
Data Examples, Quality Control Data & Sample Dilution
Validation Standard Curve O.D. At 450nm
Concentration (pg/ml) | 0 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 | 2000 |
O.D. | 0.020 | 0.055 | 0.113 | 0.189 | 0.345 | 0.589 | 1.106 | 1.913 |
Data Example Images
Click image to see more details
Human VEGF PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect VEGFA, Dilution ratio of 1:1, concentration in serum and plasma is around 80 pg/ml..
Some articles we found to cite concentrations of VEGFA in samples: 25374209 (Pubmed IDs).
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human VEGF ELISA Kit PicoKine® within the same batch/lot to ensure the consistency of the kits' performances. ELISA intra assay consistency is measured using wells from the same plate/assay kit. ELISA inter assay consistency is measured using wells from different plates from the same batch production/lot.
Intra-Assay Precision | Inter-Assay Precision | |||||
---|---|---|---|---|---|---|
Sample | 1 | 2 | 3 | 1 | 2 | 3 |
n | 16 | 16 | 16 | 24 | 24 | 24 |
Mean (pg/ml) | 72 | 270 | 946 | 76 | 254 | 955 |
Standard deviation | 4.53 | 21.33 | 59.59 | 9.62 | 24.38 | 74.49 |
CV (%) | 9.6% | 7.9% | 9.6% | 7.8% | 9.6% | 7.8% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of VEGFA in ELISA kits from four different production batches/lots.
Lots | Lot 1 (pg/ml) | Lot 2 (pg/ml) | Lot 3 (pg/ml) | Lot 4 (pg/ml) | Mean (pg/ml) | Standard Deviation | CV (%) |
---|---|---|---|---|---|---|---|
Sample 1 | 72 | 80 | 82 | 71 | 76 | 4.81 | 6.3% |
Sample 2 | 270 | 267 | 252 | 245 | 258 | 10.35 | 4% |
Sample 3 | 946 | 897 | 1026 | 1037 | 976 | 57.79 | 5.9% |
Protein Target Info & Infographic
Gene/Protein Information For VEGFA (Source: Uniprot.Org, NCBI)
Gene Name
VEGFA
Full Name
Vascular endothelial growth factor A
Weight
27042 MW
Superfamily
PDGF/VEGF growth factor family
Alternative Names
Vascular endothelial growth factor A;VEGF-A;Vascular permeability factor;VPF;VEGFA;VEGF; VEGFA MVCD1, VEGF, VPF vascular endothelial growth factor A vascular endothelial growth factor A|vascular endothelial growth factor A121|vascular endothelial growth factor A165|vascular permeability factor
*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".For more info on VEGFA, check out the VEGFA Infographic
We have 30,000+ of these available, one for each gene! check them out.
In this infographic you will see the following information for VEGFA: database IDs, super-family, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact us [email protected].
Specific Publications For Human VEGF ELISA Kit PicoKine® (EK0539)
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Customer Reviews
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Customer Q&As
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Find answers in Q&As, reviews.
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21 Customer Q&As for Human VEGF ELISA Kit PicoKine®
Question
Q: we need your suggestion regarding the dilution ratio of serum samples for detection of VEGFA in Human plasma? I am trying to measure a a number of analytes and it requires 100ul of diluted samples for each well. We have limited sample quantitys so we like to dilute as much as possible.
Verified Customer
Verified customer
Asked: 2020-10-16
Answer
A: unable to know the physiological or pathological context of your samples we cannot recommend a dilution ratio without performing a pilot test with your samples. Here is how you can perform a pilot study on your own: perform a serial dilution of your samples on the VEGFA ELISA kit to make sure you have a linear ascending curve followed by a plateau, which signifies the samples saturating the detection limit of the kit. Then you can pick the dilution ratios from samples in the linear part of the curve as your experimental dilution ratio.
If you are interested in using our ELISA service, you can also send us your sample and we will take care of everything for you. You can check our service details here: bosterbio.com/services/assay-services/ELISA-testing-service
Since you mentioned you have limited samples, our cost effective multiplex ELISA service would fit perfectly for your needs, where we can generate dozens of data points using as little as 25ul sample volume. Information on this service is also in the above link.
Boster Scientific Support
Answered: 2020-10-16
Question
Q: Can VEGFA ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
Verified Customer
Verified customer
Asked: 2020-08-25
Answer
A: Unfortunately, Boster Bio has not routinely validated tissue homogenates as a sample type for ELISA kits. This does not mean that ELISA kits are not valid for other sample types than we have tested: it means further investigation is needed. One will need to perform a spike and recovery study to determine if an unvalidated sample type will work with a particular kit. To perform a spike and recovery experiment, one should divide a sample into two aliquots. In one of the aliquots, the user should spike in a known amount of the kit standard. a dilution series is performed comparing the spiked versus the unspiked sample. Generally, samples with expected recovery and linearity between 80-120% are considered acceptable. This method can be used to validate any sample type that has not been evaluated by Boster Bio. for a more detailed spike and recovery protocol, please contact technical support.
Note: acceptable ranges should be determined individually by each laboratory. Additionally, technical support can help determine if a buffer component is not compatible with a given ELISA kit. please check the Citations tab on the product webpage for peer-reviewed papers utilizing a wide range of sample types. We also have an innovator's reward program where if the user validates our ELISA kits in applications or samples previously not validated by Boster Bio or other users, and share such information with us by submit a review, we will reward the user's efforts with a free antibody or ELISA kit from our catalog. Biocompare.com will also give $20 Amazon giftcard as an additional reward, if the review is submitted there as well.
Boster Scientific Support
Answered: 2020-08-25
Question
Q: how much samples can be assayed in a Picokine® ELISA Kit?
Z. Jackson
Verified customer
Asked: 2020-07-22
Answer
A: The Picokine® ELISA Kits will generally run a 7-point standard curve, non-specific binding wells, and 39 samples in duplicate. this may depends upon the kit used so please refer to each datasheet for details.
Boster Scientific Support
Answered: 2020-07-22
Question
I would like to know the epitopes and cross reactivity of this Human VEGF ELISA Kit PicoKine antibody as well as the animal of origin.
Verified Customer
Verified customer
Asked: 2020-03-23
Answer
The Human VEGF ELISA Kit PicoKine, Capture antibody; monoclonal mouse and the immunogen sequence is Ala27
Boster Scientific Support
Answered: 2020-03-23
Question
Q: how to analyze ELISA data? I measured VEGFA level in plasma.
Verified Customer
Verified customer
Asked: 2020-02-09
Answer
A: please read this article on ELISA data analysis. bosterbio.com/ELISA-data-analysis-instructions. we also provide a convenient online tool that you can use to analyze ELISA data. bosterbio.com/biology-research-tools/ELISA-data-analysis-online
Boster Scientific Support
Answered: 2020-02-09
Question
I want to measure VEGF in urine using ELISA. I saw your product Human VEGF ELISA Kit PicoKine ,is it suitable for urine samples?
Verified Customer
Verified customer
Asked: 2020-01-16
Answer
The Human VEGF ELISA Kit PicoKine is suitable in detecting VEGF in biological fluids such as cell culture supernatant, serum and plasma.
Boster Scientific Support
Answered: 2020-01-16
Question
Has your lab ever tested EK0539 in urine samples? Or, have you had a customer that has had success/failure in testing this kit with urine samples?
Verified Customer
Verified customer
Asked: 2019-11-25
Answer
Our lab technicians haven't
Boster Scientific Support
Answered: 2019-11-25
Question
What is being used in your method when transfering genes to sf21 cells for EK0539?
Verified customer
Asked: 2019-11-14
Answer
For the Human VEGF ELISA Kit PicoKine™ (EK0539), we use baculovirus to transfer genes to sf21 cells.
Boster Scientific Support
Answered: 2019-11-14
Question
How come I saw no signal for running your kit with my VEGF plasma sample?
Verified Customer
Verified customer
Asked: 2019-11-13
Answer
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.
Boster Scientific Support
Answered: 2019-11-13
Question
What is a possible cause for obtaining low data results with EK0539?
Verified customer
Asked: 2019-11-11
Answer
For the Human VEGF ELISA Kit PicoKine® (EK0539), the lower data might be caused by some factors which include but not limited to unstable temperature during transportation and variation of operation conditions.
Boster Scientific Support
Answered: 2019-11-18
Question
Have other isoforms been tested for cross-reactivity with EK0541 and EK0539?
Verified customer
Asked: 2019-09-25
Answer
The The Human VEGF ELISA Kit PicoKine EK0539 and Mouse VEGF ELISA Kit PicoKine EK0541 are cross-reactive with their specific targets only.
Boster Scientific Support
Answered: 2019-09-26
Question
Q: Are Boster Bio recombinant proteins and antibodies sterile?
P. Dayal
Verified customer
Asked: 2019-08-10
Answer
A: although the vials are bottled using aseptic techniques, heat-treated vials, and sterile stock solutions, they are not considered or guaranteed to be sterile. If sterile material is needed for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.
Boster Scientific Support
Answered: 2019-08-10
Question
Q: how long can samples (cell cultures, serum, and plasma) be stored and still be stable for quantification VEGFA using the EK0539 Human VEGFA Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2018-09-30
Answer
A: Boster Bio does not assess sample stability. Variations in sample collection, processing, and storage may alter the stabilityof samples. It is recommend to assay sample right after collection when possible, or aliquot into single use volumes and store samples frozen. avoid repetitive freeze-thaw cycles with the stored samples in order to prevent protein degradation.
Boster Scientific Support
Answered: 2018-09-30
Question
Q: how are cell lysates prepared for use in Picokine® ELISA kits?
Verified Customer
Verified customer
Asked: 2018-09-29
Answer
A: in those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate can be found in the product insert. Components in lysate and lysis buffer can affect immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.
Boster Scientific Support
Answered: 2018-09-29
Question
Q: how to thaw whole blood sample for VEGFA ELISA after freezing?
Verified Customer
Verified customer
Asked: 2018-09-12
Answer
A: we do not recommend freezing and thaw whole blood. erythrocytes are fragile and, if frozen and thawed, will undergo hemolysis rendering the samples useless. To keep your blood samples to test VEGFA for a later time, you should let the blood clot in glass tubes and collect the serum to freeze for later use.
Boster Scientific Support
Answered: 2018-09-12
Question
Q: is there any online tool I can use to streamline the data analysis for my ELISA results?
Verified Customer
Verified customer
Asked: 2018-06-26
Answer
A: We have a web based ELISA curve fitting (4pl) and data analysis tool. Please give it a try: bosterbio.com/biology-research-tools/ELISA-data-analysis-online. You can also consult our article on ELISA data analysis: bosterbio.com/ELISA-data-analysis-instructions
Boster Scientific Support
Answered: 2018-06-26
Question
Q: if the enzyme conjugated VEGFA antibodies are mixed with the substrate, will that change the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the VEGFA antigen?
Verified Customer
Verified customer
Asked: 2018-03-23
Answer
A: The enzyme is always active. Avoid contaminating the substrate with enzyme prior to the incubation otherwise it compromises the assay with false positive signal.
Boster Scientific Support
Answered: 2018-03-23
Question
Does this kit detect all forms of VEGF or just VEGFA
Verified Customer
Verified customer
Asked: 2017-08-02
Answer
This assay detects VEGFA
Boster Scientific Support
Answered: 2017-08-02
Question
I would like to purchase Human VEGF ELISA Kit PicoKine and use it on protein extracts in lysis buffer containing 8M urea. Do you have any information if the ELISA work with Urea 1M after dilution of the sample in diluent buffer?
Verified Customer
Verified customer
Asked: 2017-05-12
Answer
Unfortunately, we have not tested the effects of Urea containing lysis buffer on samples with this antibody Human VEGF ELISA Kit PicoKine, so we do not have any idea how well this would work. But it is important to note that Urea is a Chaotropic agent at 6 - 8 M that means it can denature proteins, thus this buffer formula may unfavorably affect the VEGF protein causing it not to be recognized by the ELISA.
Boster Scientific Support
Answered: 2017-05-12
Question
Can you tell me how to troubleshoot?
Verified Customer
Verified customer
Asked: 2016-09-15
Answer
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.
Boster Scientific Support
Answered: 2016-09-15
Question
For the kit EK0539 a customer ask if the kit detects all forms of VEGF or only VEGFA?
Verified Customer
Verified customer
Asked: 2014-02-10
Answer
EK0539 detects just VEGFA.
Boster Scientific Support
Answered: 2014-02-10