Human E-Cad (E-Cadherin) CLIA Kit

  • CLIA-Elabscience
  • CLIA-Elabscience
  • CLIA-Elabscience
    • CLIA-Elabscience
    • CLIA-Elabscience
    • CLIA-Elabscience
    <
    >
    • CLIA-Elabscience
    • CLIA-Elabscience
    • CLIA-Elabscience
    • CLIA-Elabscience
    • CLIA-Elabscience
    • CLIA-Elabscience
      Catalog number:E-CL-H0014

      Synonyms:CDH1, Arc-1, CD324, CDHE, LCAM, UVO, CAM 120/80, Epithelial Cadherin, Uvomorulin

      Size:
      • 96T
      Qty:
      - +
      Price: Inquire

      Reactivity: Human

      Detection Range: 31.25~2000 pg/mL

      Sensitivity: 18.75 pg/mL

      Lead Time: 10-12daysWelcome to order from local distributors.

      Add to cart Compare Bulk request Manual

      Intended use

      This CLIA kit can be applied to the in vitro quantitative determination of Human E-Cad concentrations in serum, plasma and other biological fluids.

      Test principle

      This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human E-Cad. Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human E-Cad and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human E-Cad, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human E-Cad. The concentration of Human E-Cad in the samples can be calculated by comparing the RLU of the samples to the standard curve.

      Assay type Sandwich
      Format 96T
      Assay time 4.5h
      Reactivity Human
      Detection method Chemiluminescence
      Detection range 31.25-2000 pg/mL
      Sensitivity 18.75 pg/mL
      Sample volume 100μL
      Sample type Serum, plasma and other biological fluids
      Repeatability CV < 15%

      Specificity

      This kit recognizes natural and some recombinant Human E-Cad. No significant cross-reactivity or interference between Human E-Cad and analogues 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) RLU Average Corrected
      2000 53213
      58229
      55721 55693
      1000 23322
      24626
      23974 23946
      500 11531
      10533
      11032 11004
      250 5144
      5444
      5294 5266
      125 2664
      2552
      2608 2580
      62.5 1336
      1286
      1311 1283
      31.25 665
      683
      674 646
      0 27
      29
      28 --

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Human E-Cad were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Human E-Cad 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)
      97.29 314.83 843.78 97.29 314.83 843.78
      Standard
      deviation
      8.35 34.41 76.36 8.35 34.41 76.36
      C V (%) 8.58 10.93 9.05 8.58 10.93 9.05

      Recovery

      The recovery of Human E-Cad 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) 89-102 95
      EDTA plasma (n=5) 94-107 100
      Cell culture media (n=5) 96-110 104

      Linearity

      Samples were spiked with high concentrations of Human E-Cad 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 (%) 89-103 95-110 89-101
      Average (%) 95 101 94
      1:4 Range (%) 88-102 85-97 88-100
      Average (%) 95 92 94
      1:8 Range (%) 90-106 99-115 103-118
      Average (%) 97 105 109
      1:16 Range (%) 86-99 101-114 96-110
      Average (%) 92 107 102

      Kit Components& Storage

      The unopened kit can be stored at 4 ℃ for 1 week. If the kit is not used within 1 week, store the items separately according to the following conditions since the kit is received.

      Item Specifications Storage
      Micro CLIA Plate 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
      Plate Sealer 5 pieces
      Product Description 1 copy
      Certificate of Analysis 1 copy

      Note:
      All reagent bottle cap must be tighten 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

      • Chemiluminescence immunoassay analyzer
      • 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 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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