Human C4 (Complement Component 4) CLIA Kit

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

      Catalog number:E-CL-H0154

      Synonyms:C4

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

      Reactivity: Human

      Detection Range: 93.75~6000 pg/mL

      Sensitivity: 56.25 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 C4 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 C4. 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 C4 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 C4, 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 C4. You can calculate the concentration of Human C4 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 93.75-6000 pg/mL
      Sensitivity 56.25 pg/mL
      Sample volume 100μL
      Sample type Serum, plasma and other biological fluids
      Repeatability CV < 15%

      Specificity

      This kit recognizes Human C4 in samples. No significant cross-reactivity or interference between Human C4 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
      6000 59656
      71610
      65633 65614
      3000 33916
      36062
      34989 34970
      1500 18164
      17344
      17754 17735
      750 8031
      9283
      8657 8638
      375 4267
      3711
      3989 3970
      187.5 1712
      1538
      1625 1606
      93.75 417
      455
      436 417
      0 18
      20
      19 --

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human C4 were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human C4 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) 329.42 570.42 2771.77 338.58 604.86 2670.80
      Standard deviation 31.29 67.82 167.97 27.26 71.25 198.17
      C V (%) 9.50 11.89 6.06 8.05 11.78 7.42

      Recovery

      The recovery of Human C4 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) 93-108 100
      EDTA plasma (n=5) 101-113 108
      Cell culture media (n=5) 100-115 107

      Linearity

      Samples were spiked with high concentrations of Human C4 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 (%) 102-116 93-107 88-102
      Average (%) 108 100 94
      1:4 Range (%) 95-111 97-112 102-114
      Average (%) 102 105 108
      1:8 Range (%) 89-105 101-118 87-100
      Average (%) 97 110 93
      1:16 Range (%) 96-108 100-114 97-110
      Average (%) 102 107 103

      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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