Human CASP3 (Caspase 3) CLIA Kit

  • CLIA-Elabscience
  • CLIA-Elabscience
  • CLIA-Elabscience
    • CLIA-Elabscience
    • CLIA-Elabscience
    • CLIA-Elabscience
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    • CLIA-Elabscience
    • CLIA-Elabscience
    • CLIA-Elabscience
    • CLIA-Elabscience
    • CLIA-Elabscience
    • CLIA-Elabscience
      Catalog number:E-CL-H0017

      Synonyms:CPP32, CPP32B, SCA-1, Apoptain, Yama

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

      Reactivity: Human

      Detection Range: 78.13~5000 pg/mL

      Sensitivity: 46.88 pg/mL

      Lead Time: 7~10 days

      Add to cart Compare Bulk request Manual

      Order directly online from USA, please contact us firstly if order from the other countries.

      Intended use

      This CLIA kit can be applied to the in vitro quantitative determination of Human CASP3 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 CASP3. 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 CASP3 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 CASP3, 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 CASP3. The concentration of Human CASP3 in the samples can be calculated by comparing the OD of the samples to the standard curve.

      Assay type Chemiluminescent-ELISA
      Format 96T
      Assay time 4.5h
      Reactivity Human
      Detection method Chemiluminescen
      Detection range 78.13-5000 pg/mL
      Sensitivity 46.88 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 CASP3. No significant cross-reactivity or interference between Human CASP3 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
      5000 62415
      67873
      65144 65114
      2500 32539
      36675
      34607 34577
      1250 19643
      17135
      18389 18359
      625 9793
      10085
      9939 9909
      312.5 5593
      4773
      5183 5153
      156.25 2694
      2578
      2636 2606
      78.13 1337
      1353
      1345 1315
      0 29
      31
      30 --

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Human CASP3 were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Human CASP3 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)
      238.20 766.53 1722.76 238.20 766.53 1722.76
      Standard
      deviation
      26.96 54.35 193.47 26.96 54.35 193.47
      C V (%) 11.32 7.09 11.23 11.32 7.09 11.23

      Recovery

      The recovery of Human CASP3 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) 84-99 91
      EDTA plasma (n=5) 87-101 94
      Cell culture media (n=5) 98-110 104

      Linearity

      Samples were spiked with high concentrations of Human CASP3 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 (%) 95-109 103-120 93-108
      Average (%) 101 110 100
      1:4 Range (%) 88-100 95-109 86-99
      Average (%) 94 101 92
      1:8 Range (%) 88-98 98-113 92-104
      Average (%) 93 104 97
      1:16 Range (%) 92-107 102-114 96-112
      Average (%) 100 108 102

      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, 10 mL
      HRP Conjugate Diluent 1 vial, 10 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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