Human RANTES (Regulated On Activation, Normal T-Cell Expressed and Secreted) 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-H0023

      Synonyms:CCL5, SCYA5, SISd, TCP228, D17S136E

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

      Reactivity: Human

      Detection Range: 15.63~1000 pg/mL

      Sensitivity: 9.38 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 RANTES 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 RANTES. 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 RANTES 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 RANTES, 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 RANTES. The concentration of Human RANTES 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 15.63-1000 pg/mL
      Sensitivity 9.38 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 RANTES. No significant cross-reactivity or interference between Human RANTES 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
      1000 50664
      60148
      55406 55378
      500 22370
      26956
      24663 24635
      250 11226
      10026
      10626 10598
      125 4859
      5161
      5010 4982
      62.5 2650
      2444
      2547 2519
      31.25 1438
      1262
      1350 1322
      15.63 722
      808
      765 737
      0 27
      29
      28 --

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Human RANTES were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Human RANTES 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)
      46.85 110.70 377.51 46.85 110.70 377.51
      Standard
      deviation
      5.95 13.15 30.47 5.95 13.15 30.47
      C V (%) 12.70 11.88 8.07 12.70 11.88 8.07

      Recovery

      The recovery of Human RANTES 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) 89-103 95
      EDTA plasma (n=5) 87-101 92
      Cell culture media (n=5) 87-100 92

      Linearity

      Samples were spiked with high concentrations of Human RANTES 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 (%) 100-116 101-117 101-114
      Average (%) 106 109 107
      1:4 Range (%) 92-104 102-119 86-100
      Average (%) 99 109 93
      1:8 Range (%) 94-108 93-110 96-112
      Average (%) 101 100 103
      1:16 Range (%) 98-113 97-114 99-110
      Average (%) 106 105 105

      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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