Human COL7 (Collagen Type Ⅶ) CLIA Kit

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

      Catalog number:E-CL-H0561

      Size:
      • 96T
      Qty:
      - +
      Price: $682

      Reactivity: Human

      Detection Range: 31.25~2000 pg/mL

      Sensitivity: 18.75 pg/mL

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

      Add to cart Compare Bulk request

      Intended use

      This CLIA kit applies to the in vitro quantitative determination of Human COL7 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 COL7. 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 COL7 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 COL7, 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 COL7. You can calculate the concentration of Human COL7 in the samples by comparing the RLU value of the samples to the standard curve.

      Assay type Sandwich
      Format 96T
      Assay time 3.5h
      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 Human COL7 in samples. No significant cross-reactivity or interference between Human COL7 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
      2000 52193
      59249
      55721 55693
      1000 22505
      25443
      23974 23946
      500 11496
      10568
      11032 11004
      250 4898
      5690
      5294 5266
      125 2750
      2466
      2608 2580
      62.5 1366
      1256
      1311 1283
      31.25 662
      686
      674 646
      0 27
      29
      28 --

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human COL7 were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human COL7 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) 99.67 298.14 708.17 105.25 327.56 674.86
      Standard deviation 10.81 21.14 68.48 12.86 29.68 51.36
      C V (%) 10.85 7.09 9.67 12.22 9.06 7.61

      Recovery

      The recovery of Human COL7 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) 87-101 92
      EDTA plasma (n=5) 98-114 105
      Cell culture media (n=5) 85-101 92

      Linearity

      Samples were spiked with high concentrations of Human COL7 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 (%) 99-112 87-99 96-109
      Average (%) 107 94 102
      1:4 Range (%) 84-97 95-107 95-110
      Average (%) 90 101 102
      1:8 Range (%) 94-110 93-107 96-108
      Average (%) 100 99 101
      1:16 Range (%) 99-115 88-102 91-105
      Average (%) 106 93 98

      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 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 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 A 1 vial, 5 mL 2-8℃ (shading light)
      Substrate Reagent B 1 vial, 5 mL 2-8℃ (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

      elisa assay procedure 1

      1.Add 100 μL of standard or sample to each well. Incubate for 90 min at 37℃.

      elisa assay procedure 2

      2.Remove the liquid.

      elisa assay procedure 3

      3.Add 100 μL Biotinylated Detection Ab. Incubate 1 hour at 37℃. Aspirate and wash 3 times.

      elisa assay procedure 4

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

      elisa assay procedure 5

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

      elisa assay procedure 6

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

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