Rat HSPG (Heparan Sulphate Proteoglycan) CLIA Kit

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

      Catalog number:E-CL-R0336

      Synonyms:SDC2, Syndecan-2, CD362, HSPG1, SYND2

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

      Reactivity: Rat

      Detection Range: 62.50~4000 pg/mL

      Sensitivity: 37.50 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 Rat HSPG 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 Rat HSPG. Standards or samples are added to the micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat HSPG 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 Rat HSPG, 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 Rat HSPG. You can calculate the concentration of Rat HSPG 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 Rat
      Detection method Chemiluminescence
      Detection range 62.50-4000 pg/mL
      Sensitivity 37.50 pg/mL
      Sample volume 100μL
      Sample type Tissue homogenates
      Repeatability CV < 15%

      Specificity

      This kit recognizes Rat HSPG in samples. No significant cross-reactivity or interference between Rat HSPG 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
      4000 57272
      60890
      59081 59046
      2000 25760
      29958
      27859 27824
      1000 13736
      13232
      13484 13449
      500 6418
      6792
      6605 6570
      250 3390
      3096
      3243 3208
      125 1608
      1554
      1581 1546
      62.50 718
      792
      755 720
      0 34
      36
      35 --

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat HSPG were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat HSPG 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) 193.74 506.22 1831.97 176.71 544.75 1979.60
      Standard deviation 23.38 57.00 128.42 21.51 52.90 192.02
      C V (%) 12.07 11.26 7.01 12.17 9.71 9.70

      Recovery

      The recovery of Rat HSPG 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) 97-112 102
      EDTA plasma (n=5) 92-106 98
      Cell culture media (n=5) 89-101 94

      Linearity

      Samples were spiked with high concentrations of Rat HSPG 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 (%) 89-101 99-114 102-117
      Average (%) 96 108 108
      1:4 Range (%) 85-100 89-100 99-115
      Average (%) 92 94 105
      1:8 Range (%) 97-110 88-104 100-112
      Average (%) 104 95 106
      1:16 Range (%) 103-116 91-105 98-113
      Average (%) 109 97 106

      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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