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Human VEGFR-2/KDR(Vascular Endothelial Growth Factor Receptor 2) ELISA Kit

Citations(9)Uniprot : P35968

Cat.No.:E-EL-H1603
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	78.13-5000 pg/mL
Sensitivity	46.88 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human VEGFR-2/KDR in samples. No significant cross-reactivity or interference between Human VEGFR-2/KDR and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Human VEGFR-2/KDR concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human VEGFR-2/KDR. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human VEGFR-2/KDR and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human VEGFR-2/KDR, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human VEGFR-2/KDR. You can calculate the concentration of Human VEGFR-2/KDR in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human VEGFR-2/KDR were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human VEGFR-2/KDR were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	243.80	712.30	2220.00	232.10	733.70	2091.00
Standard deviation	14.90	30.60	119.90	12.30	40.40	102.50
CV (%)	6.11	4.30	5.40	5.30	5.51	4.90

Recovery

The recovery of Human VEGFR-2/KDR spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	91-105	97
EDTA plasma (n=8)	93-109	101
Cell culture media (n=8)	88-103	94

Linearity

Samples were spiked with high concentrations of Human VEGFR-2/KDR and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P35968
Synonyms	VEGFR2, CD309, FLK1
Research Area	Cancer, Cardiovascular, Metabolism, Microbiology, Signal transduction, Stem cells

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

Arabian Journal of Chemistry (2021) IF: 5.165
Inhibition effects of 7-phloro-eckol from Ecklonia cava on metastasis and angiogenesis induced by hypoxia through regulation of AKT/mTOR and ERK signaling pathways

DOI: 10.1016/j.arabjc.2021.103187

Sample: Cell lysate
BIOORGANIC CHEMISTRY (2021) IF: 5.275
Pharmacophore based drug design and synthesis of oxindole bearing hybrid as anticancer agents

DOI: 10.1016/j.bioorg.2021.105358

PMID: 34544029
Sample: A431 cell
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY (2021) IF: 5.279
Mechanism Analysis of Antiangiogenic d-Isofloridoside from Marine Edible Red algae Laurencia undulata in HUVEC and HT1080 cell

DOI: 10.1021/acs.jafc.1c05007

PMID: 34767715
Sample: HT1080 cells,HUVEC cells
Chinese Journal of Cancer Research (2020) IF: 4.135
Tim-3 promotes cell aggressiveness and paclitaxel resistance through NF-κB/STAT3 signalling pathway in breast cancer cells

DOI: 10.21147/j.issn.1000-9604.2020.05.02

PMID: 33223752
Sample: Cell culture supernatant
INTERNATIONAL IMMUNOPHARMACOLOGY (2022) IF: 4.932
Mechanism of two alkaloids isolated from coral endophytic fungus for suppressing angiogenesis in atherosclerotic plaque in HUVEC

DOI: 10.1016/j.intimp.2022.108931

PMID: 35704971
Sample: HUVEC
JOURNAL OF ETHNOPHARMACOLOGY (2020) IF: 3.69
Anti-angiogenic effect of Shikonin in rheumatoid arthritis by downregulating PI3K/AKT and MAPKs signaling pathways

DOI: 10.1016/j.jep.2020.113039

PMID: 32497675
Sample: Serum,Cell culture supernatant
JOURNAL OF MOLECULAR STRUCTURE (2020) IF: 2.463
Synthesis, crystal structure, cytotoxicity and evaluation of the 6-oxo-6H-pyrrolo[3,2,1-ij]quinoline-5-carbaldehydes for inhibitory effect against protein kinases (VEGFR-2 and EGFR) and cyclooxygenase-2 (COX-2) activities

DOI: 10.1016/j.molstruc.2020.128907

Sample: MDA-MB-231 cell
BIOSCIENCE REPORTS (2020) IF: 2.942
Tim-3 promotes tube formation and decreases tight junction formation in vascular endothelial cells

DOI: 10.1042/BSR20202130

PMID: 33015716
Sample: HMVECs,HUVECs
MEDICAL SCIENCE MONITOR (2019) IF: 1.98
Serum Levels of Angiogenic Factors Distinguish Between Women with Preeclampsia and Normotensive Pregnant Women But Not Severity of Preeclampsia in an Obstetric Center in Turkey

DOI: 10.12659/MSM.915092

PMID: 31522187
Sample: Serum

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Reviews/Q&A

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Reviews
Q&A

M***************lSubmitted [ Jul 14 2022 ]

Asked: what is the normal range of Human VEGF R2/KDR in pg/mL ?

adminSubmitted [ Jul 14 2022 ]

Answered: For the kit E-EL-H1603, the reference value of our internal human serum/plasma sample is 16-27ng/mL, and it is recommended to dilute 50-100 times for detection.

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