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SKU:EK0544
Application:ELISA
Sensitivity:<4pg/ml
Sample Type:cell culture supernates, cell lysates, serum and plasma (heparin, EDTA).
Price: $400.00
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Overview

Product Name Human VEGFR2/KDR PicoKine™ ELISA Kit
Description Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human VEGFR2/KDR. 96wells/kit, with removable strips.
Cite This Product Human VEGFR2/KDR PicoKine™ ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0544)
Validated Species Human
Application ELISA

*Our Boster Guarantee covers the use of this product in the above tested applications.

**For positive and negative control design, consult "Tissue specificity" under Protein Target Info.

Immunogen Expression system for standard: NSO; Immunogen sequence: A20-E764
Cross Reactivity There is no detectable cross-reactivity with other relevant proteins.
Pack Size 96wells/kit, with removable strips.

Properties

Sensitivity <4pg/ml
*Sensitivity, or Lower Limit of Detection (LLD), is the minimum level of target protein the ELISA assay can detect. We measure 20 blank wells and if the O.D. value is 2 standard deviations higher than the blanks' average O.D. the sample can be deemed positive.
Assay Range 62.5pg/ml-4000pg/ml
*This assay range is determined using common samples. For samples with low target protein concentrations, users can adjust the standard curve to extend the lower limit of assay range.
Sample Type cell culture supernates, cell lysates, serum and plasma (heparin, EDTA).

*The above listed samples are the ones valided with the assay. If you do not see your sample of interest listed, as long as there is enough level of target protein present in the sample, this Picokine™ ELISA kit should detect it.
**For protocol and tips regarding preparing your sample of interest, please check our ELISA sample preparation guide.
Capture Antibody monoclonal antibody from mouse
Detection Antibody polyclonal antibody from goat
Natural Levels Some research articles suggesting the natural levels of KDR are listed below:
Storage Store at 4˚C for 6 months, at -20˚C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.)

Kit Components

Catalog numberDescriptionQuantity
EK0544-CAP96-well plate precoated with anti-Human KDR antibody1
EK0544-STlyophilized recombinant Human KDR standard10ng/tubex2
EK0544-DAbiotinylated anti-Human KDR antibody130ul(dilution 1:100)
AR1103Avidin-Biotin-Peroxidase Complex(ABC)130ul(dilution 1:100)
AR1106-1sample diluent buffer30ml
AR1106-2antibody diluent buffer12ml
AR1106-3ABC diluent buffer12ml
AR1104TMB color developing agent10ml
AR1105TMB stop solution10ml
PLA-SEAAdhesive cover4
AR0030-EPBS washing bufferPowder for 1000ml

Washing buffer Preparation: Disolve AR0030-E to 1000ml distilled water and adjust pH to 7.2~7.6. Finally, adjust the total volume to 1L.

*Additional components can be purchased. If you need extra of the above components please order them together to avoid additional shipping charges.

See Prices For Extra Components

Material Required But Not Provided

  • 1. Microplate reader in standard size.
  • 2. Automated plate washer.
  • 3. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
  • 4. Clean tubes and Eppendorf tubes.
*Reagents needed for sample collection and preparation are not included in the kit.

Protein Target Info (Source: Uniprot.org)

You can check the tissue specificity below for information on selecting positive and negative control.

Gene Name KDR
Protein Name Vascular endothelial growth factor receptor 2
Molecular Weight 151527 MW
Protein Function Tyrosine-protein kinase that acts as a cell-surface receptor for VEGFA, VEGFC and VEGFD. Plays an essential role in the regulation of angiogenesis, vascular development, vascular permeability, and embryonic hematopoiesis. Promotes proliferation, survival, migration and differentiation of endothelial cells. Promotes reorganization of the actin cytoskeleton. Isoforms lacking a transmembrane domain, such as isoform 2 and isoform 3, may function as decoy receptors for VEGFA, VEGFC and/or VEGFD. Isoform 2 plays an important role as negative regulator of VEGFA- and VEGFC-mediated lymphangiogenesis by limiting the amount of free VEGFA and/or VEGFC and preventing their binding to FLT4. Modulates FLT1 and FLT4 signaling by forming heterodimers. Binding of vascular growth factors to isoform 1 leads to the activation of several signaling cascades. Activation of PLCG1 leads to the production of the cellular signaling molecules diacylglycerol and inositol 1,4,5-trisphosphate and the activation of protein kinase C. Mediates activation of MAPK1/ERK2, MAPK3/ERK1 and the MAP kinase signaling pathway, as well as of the AKT1 signaling pathway. Mediates phosphorylation of PIK3R1, the regulatory subunit of phosphatidylinositol 3-kinase, reorganization of the actin cytoskeleton and activation of PTK2/FAK1. Required for VEGFA-mediated induction of NOS2 and NOS3, leading to the production of the signaling molecule nitric oxide (NO) by endothelial cells. Phosphorylates PLCG1. Promotes phosphorylation of FYN, NCK1, NOS3, PIK3R1, PTK2/FAK1 and SRC. .
Tissue Specificity Detected in cornea (at protein level). Widely expressed. .
Sequence Similarities Belongs to the protein kinase superfamily. Tyr protein kinase family. CSF-1/PDGF receptor subfamily.
Subcellular Localization Cell junction . Endoplasmic reticulum . Localized with RAP1A at cell-cell junctions (By similarity). Colocalizes with ERN1 and XBP1 in the endoplasmic reticulum in endothelial cells in a vascular endothelial growth factor (VEGF)-dependent manner (PubMed:23529610). .
Uniprot ID P35968
Alternative Names Vascular endothelial growth factor receptor 2;VEGFR-2;2.7.10.1;Fetal liver kinase 1;FLK-1;Kinase insert domain receptor;KDR;Protein-tyrosine kinase receptor flk-1;CD309;KDR;FLK1, VEGFR2;
Research Areas |cardiovascular|angiogenesis|growth factors|vegf|vegf receptors| microbiology|interspecies interaction|host virus interaction| signal transduction|protein phosphorylation|tyrosine kinases|receptor tyrosine kinases|growth factors/hormones| stem cells|hematopoietic progenitors|surface molecules| cancer|invasion/microenvironment|angiogenic growth factors|oncoproteins/suppressors|oncoproteins|growth factor receptors|endothelial progenitors|endothelial markers|cancer metabolism|response to hypoxia| cardiovascular|cardiovascular markers|cell markers|endothelial cells| metabolism|pathways and processes|metabolism processes|
*if product is indicated to react with multiple species, protein info is based on the human gene.

Background for Vascular endothelial growth factor receptor 2

Vascular endothelial growth factor receptor 2(VEGFR-2) also known as Kinase insert domain receptor(KDR, a type III receptor tyrosine kinase) is a VEGF receptor. Through in situ hybridization of a genomic DNA probe to metaphase chromosomes, VEGFR2 was localized to 4q11-->q12. VEGF receptor 2 and the adherens junction act as shear-stress cotransducers, mediating the transduction of shear-stress signals into vascular endothelial cells.

Human VEGFR2/KDR PicoKine™ ELISA Kit Images

Click the images to enlarge.

Human VEGFR2/KDR PicoKine™ ELISA Kit
Human VEGFR2/KDR PicoKine ELISA Kit standard curve

Intra/Inter Assay Precision

>
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n161616242424
Mean(pg/ml)1971466245823416302863
Standard deviation9.6551.31127.817.55114.1177.5
CV(%)4.93.55.27.576.2

Typical Data Obtained from Human VEGFR2/KDR PicoKine™ ELISA Kit

(TMB reaction incubate at 37°C for 20-25min)

Concentration(pg/ml)062.5125250500100020004000
O.D.0.0650.1150.1650.2650.4460.8141.5172.462

*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.

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Publications

Wang L, Liu X, Wang H, Wang S. Oncol Lett. 2013 Jul;6(1):175-180. Epub 2013 May 15. Correlation Of The Expression Of Vascular Endothelial Growth Factor And Its Receptors With Microvessel Density In Ovarian Cancer.

FAQs

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: 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: 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 techincal 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%+ indentical, there is a high chance of cross reactivity for your species of interest.