Human VIP(Vasoactive Intestinal Peptide) ELISA Kit

Uniprot : P01282
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    <
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    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience

      Catalog number:E-EL-H2155

      Synonyms:PHM27

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

      Reactivity: Human

      Detection Range: 7.81~500 pg/mL

      Sensitivity: 4.69 pg/mL

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

      Add to cart Compare Bulk request

      Intended use

      This ELISA kit applies to the in vitro quantitative determination of Human VIP concentrations in serum, plasma and other biological fluids.

      Test principle

      This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with Human VIP. During the reaction, Human VIP in the sample or standard competes with a fixed amount of Human VIP on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Human VIP. Excess conjugate and unbound sample or standard are washed from the plate, and Avidin conjugated to Horseradish Peroxidase (HRP) are added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of Human VIP in the samples is then determined by comparing the OD of the samples to the standard curve.

      Assay type Competitive
      Format 96T
      Assay time 2.0h
      Reactivity Human
      Detection Method Colormetric
      Detection Range 7.81—500 pg/mL
      Sensitivity 4.69 pg/mL
      Sample Volume 50μL
      Sample Type Serum, plasma and other biological fluids

      Specificity

      This kit recognizes Human VIP in samples. No significant cross-reactivity or interference between Human VIP 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
      500 0.316
      0.36
      0.338
      250 0.434
      0.464
      0.449
      125 0.663
      0.633
      0.648
      62.5 0.95
      0.982
      0.966
      31.25 1.401
      1.375
      1.388
      15.63 1.844
      1.816
      1.83
      7.81 2.187
      2.193
      2.19
      0 2.71
      2.714
      2.712

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human VIP were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human VIP 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) 23.50 74.30 194.50 22.70 75.30 202.10
      Standard deviation 1.50 3.60 6.80 1.20 3.80 9.70
      C V (%) 6.38 4.85 3.50 5.29 5.05 4.80

      Recovery

      The recovery of Human VIP 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) 92-104 98
      EDTA plasma (n=5) 90-103 97
      Cell culture media (n=5) 86-102 93

      Linearity

      Samples were spiked with high concentrations of Human VIP 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 (%) 89-100 93-107 92-107
      Average (%) 94 98 99
      1:4 Range (%) 83-97 92-104 99-114
      Average (%) 90 97 104
      1:8 Range (%) 87-100 93-105 97-112
      Average (%) 94 98 103
      1:16 Range (%) 83-96 86-102 95-111
      Average (%) 90 93 102

      Kit components & Storage

      An unopened kit can be stored at 2-8℃ 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 2-8℃, 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℃(shading light)
      Stop Solution 1 vial, 10 mL 2-8℃
      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

      elisa assay procedure 1

      1.Add 50 μL standard or sample to each well.

      elisa assay procedure 2

      2.Immediately add 50 μL Biotinylated Detection Ab to each well.

      elisa assay procedure 3

      3.Incubate for 45 min at 37℃. Aspirate and wash 3 times.

      elisa assay procedure 4

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

      elisa assay procedure 5

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

      elisa assay procedure 6

      6.Add 50 μL Stop Solution.

      elisa assay procedure 7

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

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