fT4(Free Thyroxine) 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-0122
      Size:
      • 96T
      • 96T*5
      Qty:
      - +
      Price: Inquire

      Reactivity: Universal

      Detection Range: 1.56~100 pg/mL

      Sensitivity: 0.94 pg/mL

      Lead Time: 3个工作日+顺丰周期Welcome to order from local distributors.

      Add to cart Compare Bulk request Manual

      Intended use

      This ELISA kit can be applied to the in vitro quantitative determination of  fT4 concentrations in serum, plasma and other biological fluids.

      Test principle

      This ELISA kit uses Competitive-ELISA as the method. The microtiter plate provided in this kit has been pre-coated with fT4. During the reaction,  fT4 in the sample or standard competes with a fixed amount of fT4 on the solid phase supporter for sites on the Biotinylated Detection Ab specific to  fT4. 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 adding Stop Solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of  fT4 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 1.56—100 pg/mL
      Sensitivity 0.94 pg/mL
      Sample Volume 50μL
      Sample Type Serum, plasma and other biological fluids

      Specificity

      This kit recognizes some recombinant and natural fT4. No significant cross-reactivity or interference was observed.

      Typical data

      For convenience in result calculation, absorbance can be used as ordinate and standard concentrations as abscissa. The standard curve provided in the manual is only for reference, and experimenters should draw the standard curve based on data of themselves.

      (pg/mL) O.D Average
      100 0.39
      0.416
      0.403
      50 0.511
      0.511
      0.511
      25 0.712
      0.696
      0.704
      12.5 1
      1.016
      1.008
      6.25 1.409
      1.385
      1.397
      3.13 1.785
      1.775
      1.78
      1.56 2.066
      2.084
      2.075
      0 2.453
      2.469
      2.461

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level fT4 were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level fT4 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)
      5.10 11.00 44.20 5.20 11.00 45.00
      Standard
      deviation
      0.30 0.50 2.30 0.30 0.50 2.00
      C V (%) 5.88 4.55 5.20 5.77 4.55 4.44

      Recovery

      The recovery of  fT4  spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.

      Sample Type Range (%) Average Recovery (%)
      Serum (n=5) 92-106 99
      EDTA plasma (n=5) 90-101 96
      Cell culture media (n=5) 96-107 102

      Linearity

      Samples were spiked with high concentrations of Human ACTH 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 (%) 87-101 99-112 91-104
      Average (%) 93 106 98
      1:4 Range (%) 85-96 92-108 100
      Average (%) 90 98 100
      1:8 Range (%) 88-104 85-98 94-109
      Average (%) 95 92 101
      1:16 Range (%) 89-106 87-99 96-111
      Average (%) 97 94 103

      Kit components & Storage

      The 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 since 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 in measuring instead of directly pouring.

      Other supplies required

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

      Assay Procedure

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

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

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

      4.Add 100 μL of HRP Conjugate to each well. 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 of Stop Solution.

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

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