Human FGF21 (Fibroblast Growth Factor 21) CLIA Kit

    • CLIA-Elabscience
    • CLIA-Elabscience
    • CLIA-Elabscience
    <
    >
    • CLIA-Elabscience
    • CLIA-Elabscience
    • CLIA-Elabscience
    • CLIA-Elabscience
    • CLIA-Elabscience
    • CLIA-Elabscience

      Catalog number:E-CL-H0074

      Synonyms:FGF-21

      Size:
      • 96T
      Qty:
      - +
      Price: $650

      Reactivity: Human

      Detection Range: 12.50~800 pg/mL

      Sensitivity: 7.50 pg/mL

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

      Add to cart Compare Bulk request Manual

      Intended use

      This CLIA kit applies to the in vitro quantitative determination of Human FGF21 concentrations in serum, plasma and other biological fluids.

      Test principle

      This CLIA kit uses the Sandwich-CLIA principle. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human FGF21. Standards or samples are added to the micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human FGF21 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 FGF21, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human FGF21. You can calculate the concentration of Human FGF21 in the samples by comparing the RLU value of the samples to the standard curve.

      Assay type Sandwich
      Format 96T
      Assay time 4.5h
      Reactivity Human
      Detection method Chemiluminescence
      Detection range 12.50-800 pg/mL
      Sensitivity 7.50 pg/mL
      Sample volume 100μL
      Sample type Serum, plasma and other biological fluids
      Repeatability CV < 15%

      Specificity

      This kit recognizes Human FGF21 in samples. No significant cross-reactivity or interference between Human FGF21 and analogues was observed.

      Typical data

      As the RLU 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)
      RLU Average Corrected
      800 48420
      56864
      52642 52616
      400 20066
      23402
      21734 21708
      200 10480
      8936
      9708 9682
      100 4366
      4738
      4552 4526
      50 2275
      2101
      2188 2162
      25 1121
      999
      1060 1034
      12.50 499
      519
      509 483
      0 25
      27
      26 --

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human FGF21 were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human FGF21 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) 39.32 81.85 297.58 39.70 80.40 316.22
      Standard deviation 4.68 7.07 23.48 3.50 7.26 28.62
      C V (%) 11.90 8.64 7.89 8.82 9.03 9.05

      Recovery

      The recovery of Human FGF21 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) 85-99 92
      EDTA plasma (n=5) 99-113 106
      Cell culture media (n=5) 97-113 104

      Linearity

      Samples were spiked with high concentrations of Human FGF21 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 (%) 95-107 97-112 88-105
      Average (%) 100 103 96
      1:4 Range (%) 101-116 96-112 95-112
      Average (%) 108 104 102
      1:8 Range (%) 98-113 103-115 83-95
      Average (%) 105 108 90
      1:16 Range (%) 90-104 95-107 87-102
      Average (%) 98 102 94

      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 CLIA 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 A 1 vial, 5 mL 4℃ (shading light)
      Substrate Reagent B 1 vial, 5 mL 4℃ (shading light)
      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

      • Chemiluminescence immunoassay analyzer
      • 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. Incubate for 90 min at 37℃.

      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 100 μL Substrate Mixture Solution. Incubate for 5 min at 37℃.

      6.Fluorescence appeared. Measure the RLU value with the Chemiluminescence immunoassay analyzer. Calculation of the results.

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