SP(Substance P) ELISA Kit

  • competitive-ELISA-Elabscience
  • competitive-ELISA-Elabscience
  • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    <
    >
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience

      Catalog number:E-EL-0067

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

      Reactivity: Universal

      Detection Range: 78.13~5000 pg/mL

      Sensitivity: 46.88 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  SP 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 SP. During the reaction,  SP in the sample or standard competes with a fixed amount of SP on the solid phase supporter for sites on the Biotinylated Detection Ab specific to  SP. 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  SP in the samples is then determined by comparing the OD of the samples to the standard curve.

      Assay type Competitive
      Format 96T
      Assay time 2.5h
      Reactivity Universal
      Detection Method Colormetric
      Detection Range 78.13—5000 pg/mL
      Sensitivity 46.88 pg/mL
      Sample Volume 50μL
      Sample Type Serum, plasma and other biological fluids

      Specificity

      This kit recognizes SP in samples. No significant cross-reactivity or interference between SP 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
      5000 0.248
      0.256
      0.252
      2500 0.383
      0.396
      0.39
      1250 0.549
      0.555
      0.552
      625 0.8149999999999999
      0.8179999999999999
      0.8169999999999999
      312.5 1.141
      1.059
      1.1
      156.25 1.401
      1.399
      1.4
      78.13 1.773
      1.695
      1.734
      0 2.378
      2.482
      2.43

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level SP were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level SP 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) 233.17 510.94 2017.45 206.77 515.17 2371.94
      Standard deviation 12.87 23.61 104.91 12.18 23.85 125.24
      C V (%) 5.52 4.62 5.20 5.89 4.63 5.28

      Recovery

      The recovery of  SP 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) 86-96 93
      EDTA plasma (n=5) 99-109 104
      Cell culture media (n=5) 90-100 96

      Linearity

      Samples were spiked with high concentrations of SP 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 (%) 93-101 94-107 90-102
      Average (%) 96 100 96
      1:4 Range (%) 97-108 86-94 94
      Average (%) 104 91 94
      1:8 Range (%) 87-95 91-103 98-112
      Average (%) 91 99 105
      1:16 Range (%) 86-97 96-108 91-103
      Average (%) 91 103 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 50 μL standard or sample to each well.

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

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

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

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

      6.Add 50 μL Stop Solution.

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

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