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

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  • sandwich-Ab-ELISA-Elabscience
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    • 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-M1292

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      Reactivity: Mouse

      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 Mouse 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 Mouse 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 Mouse 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 Mouse 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 Mouse VEGF-A. You can calculate the concentration of Mouse VEGF-A in the samples by comparing the OD of the samples to the standard curve.

      Assay type Sandwich
      Format
      Assay time
      Reactivity Mouse
      Detection Method
      Detection Range 31.25—2000 pg/mL
      Sensitivity 18.75 pg/mL
      Sample Volume Required Per Well
      Sample Type

      Specificity

      This kit recognizes Mouse VEGF-A in samples. No significant cross-reactivity or interference between Mouse 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 2.464
      2.416
      2.44 2.38
      1000 1.65
      1.611
      1.631 1.571
      500 0.967
      0.983
      0.975 0.915
      250 0.456
      0.451
      0.454 0.394
      125 0.267
      0.277
      0.272 0.212
      62.5 0.171
      0.18
      0.176 0.116
      31.25 0.111
      0.124
      0.118 0.058
      0 0.055
      0.065
      0.06 --

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse 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 Mouse 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) 94.00 212.07 852.78 83.63 230.40 893.36
      Standard deviation 4.75 10.50 46.05 4.25 11.45 47.44
      C V (%) 5.05 4.95 5.40 5.08 4.97 5.31

      Recovery

      The recovery of Mouse 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) 97-108 103
      EDTA plasma (n=5) 98-108 101
      Cell culture media (n=5) 99-109 104

      Linearity

      Samples were spiked with high concentrations of Mouse 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 (%) 86-93 82-98 92-98
      Average (%) 90 90 95
      1:4 Range (%) 92-101 87-96 92-101
      Average (%) 98 90 97
      1:8 Range (%) 90-99 91-103 88-99
      Average (%) 94 96 92
      1:16 Range (%) 91-102 89-98 91-105
      Average (%) 96 93 97

      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 of standard or sample to each well.

      2.Remove the liquid.

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

      4.Add 100 μL HRP Conjugate. Incubate 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 of Stop Solution.

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

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