QuicKey Human VEGF-A(Vascular Endothelial Growth Factor A) ELISA Kit

SKU: E-TSEL-H0026

£253⁰⁰
Save £45⁰⁰

QuicKey Human VEGF-A(Vascular Endothelial Growth Factor A) ELISA Kit

Detection Range	0.04-10 ng/mL
Sensitivity	0.03 ng/mL

Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T
Assay time	2.5h
Detection range	0.04-10 ng/mL
Sensitivity	0.03 ng/mL
Sample type &Sample volume	serum and plasma; 50μL
Specificity	This kit recognizes Human VEGF-A in samples. No significant or interference between Human VEGF-A 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 VEGF-A concentrations in serum and plasma.Please consult technical support for the applicability if other biological fluids need to be tested.

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 VEGF-A. Samples (or Standards) and biotinylated detection antibody specific for Human VEGF-A are added to the micro ELISA plate wells. Human VEGF-A would combine with the specific antibody. Then 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 VEGF-A, 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 ± 2 nm. The OD value is proportional to the concentration of Human VEGF-A. You can calculate the concentration of Human VEGF-A in the samples by comparing the OD of the samples to the standard curve.

Kit Components and Storage

An unopened kit can be stored at 2-8℃ for six months. After test, the unused wells and reagents should be stored according to the table below.

Item	Specifications	Storage conditions after test
Micro ELISA Plate (Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	2-8℃, 1 month
Reference Standard	96T: 2 vials 48T: 1 vial	Discard unused reconstituted standard and dilutions
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8℃
Biotinylated Detection Ab Working Solution	1 vial, 6 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	2-8℃ (Protect from light)
Substrate Reagent	1 vial, 10 mL	2-8℃ (Protect from light)
Stop Solution	1 vial, 10 mL	2-8℃
Plate Sealer	5 pieces
Product Description	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 VEGF-A were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human VEGF-A 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 (ng/mL)	0.48	1.13	4.28	0.42	1.19	4.51
Standard deviation	0.03	0.05	0.21	0.02	0.06	0.22
CV (%)	6.25	4.42	4.91	4.76	5.04	4.88

Recovery

The recovery of Human VEGF-A 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)	86-97	91
EDTA plasma(n=8)	82-96	90

Target Information

Background

Vascular endothelial growth factor A(VEGF-A) is a multifunctional cytokine that exerts in vivo a key role in physiological and pathological neoangiogenesis by stimulating endothelial cell proliferation and vessel hyperpermeability.[1] It is a member of the platelet-derived growth factor (PDGF)/vascular endothelial growth factor (VEGF) family and encodes a protein that is often found as a disulfide linked homodimer. In human, VEGF-A is produced by a group of isoforms as a result of alternative splicing and VEGF189, VEGF165, VEGF121 are widely expressed. Elevated levels of this protein is linked to POEMS syndrome, mutations in this gene have been associated with proliferative and nonproliferative diabetic retinopathy.[2] VEGF-A also has a major role in the pathogenesis of many other diseases including hypervascularized tumors, rheumatoid arthritis, cutaneous diseases. [3]
1. Murphy J F , Fitzgerald D J . Vascular endothelial growth factor induces cyclooxygenase-dependent proliferation of endothelial cells via the VEGF-2 receptor[J]. The FASEB Journal, 2001, 15(9):1667-1669.
2. Watanabe D, Suzuma K, Suzuma I, et al. Vitreous levels of angiopoietin 2 and vascular endothelial growth factor in patients with proliferative diabetic retinopathy[J]. American journal of ophthalmology, 2005, 139(3): 476-481.

Database Links	SwissProt: P15692 Entrez Gene: 7422
Synonyms	VEGF-A, VEGFA, MVCD1, VPF
Research Area	Cancer, Cardiovascular, Metabolism, Developmental biology, Signal transduction

Assay Procedures

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