Human PRF1 (Perforin 1/Pore-Forming Protein) CLIA Kit

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

      Catalog number:E-CL-H0749

      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 PRF1/PFP 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 PRF1/PFP. 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 PRF1/PFP 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 PRF1/PFP, 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 PRF1/PFP. You can calculate the concentration of Human PRF1/PFP 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 PRF1/PFP in samples. No significant cross-reactivity or interference between Human PRF1/PFP 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 49918
      55366
      52642 52616
      400 20487
      22981
      21734 21708
      200 10191
      9225
      9708 9682
      100 4208
      4896
      4552 4526
      50 2222
      2154
      2188 2162
      25 1100
      1020
      1060 1034
      12.50 478
      540
      509 483
      0 26
      26
      26 --

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human PRF1/PFP were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human PRF1/PFP 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) 40.88 69.68 348.80 41.60 68.44 367.31
      Standard deviation 3.73 4.91 39.03 4.69 8.12 33.79
      C V (%) 9.12 7.05 11.19 11.27 11.86 9.20

      Recovery

      The recovery of Human PRF1/PFP 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) 93-109 99
      EDTA plasma (n=5) 84-96 91
      Cell culture media (n=5) 90-104 95

      Linearity

      Samples were spiked with high concentrations of Human PRF1/PFP 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 93-106 96-111
      Average (%) 93 98 104
      1:4 Range (%) 95-111 97-112 88-103
      Average (%) 103 103 95
      1:8 Range (%) 93-107 86-101 95-111
      Average (%) 98 93 101
      1:16 Range (%) 98-110 102-116 93-109
      Average (%) 104 108 100

      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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