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

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    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
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    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience

      Catalog number:E-EL-H0111

      Synonyms:VEGF-A, VEGFA, MVCD1, VPF

      Size:
      • 96T
      • 24T
      Qty:
      - +
      Price: $495

      Reactivity: Human

      Detection Range: 31.25~2000 pg/mL

      Sensitivity: 18.75 pg/mL

      Lead Time: 7~10 daysWelcome to order from local distributors.

      Add to cart Compare Bulk request Manual

      Intended use

      This ELISA kit applies to the in vitro quantitative determination of Human VEGF-A 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 VEGF-A. 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 VEGF-A 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 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 nm ± 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.

      Assay type Sandwich
      Format 96T
      Assay time 3.5h
      Reactivity Human
      Detection Method Colormetric
      Detection Range 31.25—2000 pg/mL
      Sensitivity 18.75 pg/mL
      Sample Volume Required Per Well 100μL
      Sample Type Serum, plasma and other biological fluids

      Specificity

      This kit recognizes Human VEGF-A in samples. No significant cross-reactivity or interference between Human VEGF-A and analogues was observed.

      Typical data

      As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

      Concentration
      (pg/mL)
      O.D Average Corrected
      2000 2.491
      2.501
      2.496 2.412
      1000 1.599
      1.631
      1.615 1.531
      500 0.941
      0.905
      0.923 0.839
      250 0.426
      0.462
      0.444 0.36
      125 0.276
      0.258
      0.267 0.183
      62.5 0.199
      0.169
      0.184 0.1
      31.25 0.131
      0.141
      0.136 0.052
      0 0.081
      0.087
      0.084 --

      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 (pg/mL) 94.10 279.80 737.20 92.40 303.90 745.00
      Standard deviation 5.50 14.30 34.60 4.80 15.50 31.30
      C V (%) 5.84 5.11 4.69 5.19 5.10 4.20

      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=5) 92-105 98
      EDTA plasma (n=5) 95-110 101
      Cell culture media (n=5) 88-103 96

      Linearity

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

      Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
      1:2 Range (%) 99-114 97-111 91-105
      Average (%) 105 104 99
      1:4 Range (%) 95-112 80-95 93-106
      Average (%) 103 87 98
      1:8 Range (%) 100-116 86-100 97-110
      Average (%) 106 93 103
      1:16 Range (%) 96-110 82-92 96-107
      Average (%) 104 87 102

      Kit components & Storage

      An unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

      Item Specifications Storage
      Micro ELISA Plate(Dismountable) 8 wells ×12 strips -20℃, 6 months
      Reference Standard 2 vials
      Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 μL
      Concentrated HRP Conjugate (100×) 1 vial, 120 μL -20℃(shading light), 6 months
      Reference Standard & Sample Diluent 1 vial, 20 mL 4℃, 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 4℃(shading light)
      Stop Solution 1 vial, 10 mL 4℃
      Plate Sealer 5 pieces
      Product Description 1 copy
      Certificate of Analysis 1 copy

      Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.
      The volume of reagents in partial shipments is a little more than the volume marked on the label, please use accurate measuring equipment instead of directly pouring into the vial(s).

      Other supplies required

      • Microplate reader with 450 nm wavelength filter
      • High-precision transfer pipette, EP tubes and disposable pipette tips
      • Incubator capable of maintaining 37℃
      • Deionized or distilled water
      • Absorbent paper
      • Loading slot for Wash Buffer

      Assay Procedure

      1.Add 100 μL standard or sample to each well. Incubate for 90 min at 37℃.

      2.Remove the liquid.

      3.Add 100 μL Biotinylated Detection Ab. Incubate for 1 hour at 37℃. Aspirate and wash 3 times.

      4.Add 100 μL HRP Conjugate. Incubate for 30 min at 37℃. Aspirate and wash 5 times.

      5.Add 90 μL of Substrate Reagent. Incubate for 15 min at 37℃.

      6.Add 50 μL Stop Solution.

      7.Read at 450 nm immediately. Calculation of results.

      Citations

      1. Publication: He Y H, Li M F, Zhang X Y, et al. NLRC5 Promotes Cell Proliferation Via Regulating The AKT/VEGF-A Signaling Pathway In Hepatocellular Carcinoma[J]. Toxicology, 2016, 359: 47-57.
        Sample Type: culture supernatant
      2. Publication: Zhang F, Lu Y, Chen Q, et al. Identification Of NCK1 As A Novel Downstream Effector Of STAT3 In Colorectal Cancer Metastasis And Angiogenesis[J]. Cellular Signalling, 2017.
        Sample Type: culture supernatant
      3. Publication: Yan Y Y, Song D D, Liu L L, et al. The Relationship Of Plasma Decoy Receptor 3 And Coronary Collateral Circulation In Patients With Coronary Artery Disease[J]. Life Sciences, 2017, 189: 84-88.
        Sample Type: Plasma
      4. Publication: Liu X A, Yang Y F, Guo X J, et al. The Antiangiogenesis Effect Of Pirfenidone In Wound Healing In Vitro[J]. Journal of Ocular Pharmacology And Therapeutics, 2017, 33(9): 693-703.
        Sample Type: culture supernatant
      5. Publication: Wang J N, Yan Y Y, Song D D, et al. Reduced Plasma miR-146a Is A Predictor Of Poor Coronary Collateral Circulation In Patients With Coronary Artery Disease[J]. BioMed Research International, 2016.
        Sample Type: Plasma
      6. Publication: Jeon H, Han S R, Lee S, et al. Activation of the complement system in an osteosarcoma cell line promotes angiogenesis through enhanced production of growth factors[J]. Scientific reports, 2018, 8(1): 5415.
      7. Publication: Mesquita J, Castro-de-Sousa J P, Vaz-Pereira S, et al. Evaluation of the growth factors VEGF-a and VEGF-B in the vitreous and serum of patients with macular and retinal vascular diseases[J]. Growth Factors, 2018: 1-10.
        Sample Type: Vitreous and Serum
      8. Publication: Fei Y, Hou J, Xuan W, et al. The relationship of plasma miR-503 and coronary collateral circulation in patients with coronary artery disease[J]. Life sciences, 2018.
        Sample Type: Human
      9. Publication: L¨¹ L, Deegan A, Musa F, et al. The effects of biomimetically conjugated VEGF on osteogenesis and angiogenesis of MSCs (human and rat) and HUVECs co-culture models[J]. Colloids and Surfaces B: Biointerfaces, 2018, 167: 550-559
        Sample Type: Human
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