Human VEGF ELISA Kit PicoKine™

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The Human VEGF ELISA Kit PicoKine, Capture antibody; monoclonal mouse and the immunogen sequence is Ala27

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The Human VEGF ELISA Kit PicoKine is suitable in detecting VEGF in biological fluids such as cell culture supernatant, serum and plasma.

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Our lab technicians haven't

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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.

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This assay detects VEGFA

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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.

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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.

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EK0539 detects just VEGFA.

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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.

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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.

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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.

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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

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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.

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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.

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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.

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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.

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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

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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.

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For the Human VEGF ELISA Kit PicoKine™ (EK0539), we use baculovirus to transfer genes to sf21 cells.

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The lower data might be caused by some factors which include but not limited to unstable temperature during transportation and variation of operation conditions.

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The Human VEGF ELISA Kit PicoKine EK0539 is cross-reactive with their specific targets only.