Human GPX1(Glutathione Peroxidase 1) ELISA Kit

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  • sandwich-Ab-ELISA-Elabscience
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    • sandwich-Ab-ELISA-Elabscience
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    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
      Catalog number:E-EL-H5410
      Size:
      • 96T
      Qty:
      - +
      Price: Inquire

      Reactivity: Human

      Detection Range: 31.25~2000 pg/mL

      Sensitivity: 18.75 pg/mL

      Lead Time: 7~10 daysWelcome to order from local distributors.

      Add to cart Compare Bulk request Manual

      Intended use

      This ELISA kit can be applied to the in vitro quantitative determination of Human GPX1 concentrations in serum, plasma and other biological fluids.

      Test principle

      This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human GPX1. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human GPX1 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 GPX1, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human GPX1. The concentration of Human GPX1 in samples can be calculated by comparing the OD of the samples to the standard curve.

      Assay type Sandwich
      Format 96T
      Assay time 4.5h
      Reactivity Human
      Detection Method Colormetric
      Detection Range 31.25—2000 pg/mL
      Sensitivity 18.75 pg/mL
      Sample Volume Required Per Well 100 μL
      Sample Type Serum, plasma and other biological fluids

      Specificity

      This kit recognizes some recombinant and natural Human GPX1. No significant cross-reactivity or interference 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) O.D Average Corrected
      2000 2.521
      2.539
      2.53 2.466
      1000 1.562
      1.618
      1.59 1.526
      500 1
      0.966
      0.983 0.919
      250 0.433
      0.443
      0.438 0.374
      125 0.262
      0.24
      0.251 0.187
      62.5 0.175
      0.157
      0.166 0.102
      31.25 0.115
      0.117
      0.116 0.052
      0 0.063
      0.065
      0.064 --

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high leve Human GPX1 were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Human GPX1 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)
      110.99 204.30 946.17 118.12 214.76 1037.22
      Standard
      deviation
      5.67 8.52 51.85 6.08 9.96 44.29
      C V (%) 5.11 4.17 5.48 5.15 4.64 4.27

      Recovery

      The recovery of Human GPX1 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) 88-103 95
      EDTA plasma (n=5) 86-98 91
      Cell culture media (n=5) 86-100 91

      Linearity

      Samples were spiked with high concentrations of Human GPX1 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 (%) 97-110 97-114 96-111
      Average (%) 102 105 101
      1:4 Range (%) 91-106 79-93 88-99
      Average (%) 98 85 94
      1:8 Range (%) 90-105 85-99 88-98
      Average (%) 97 92 93
      1:16 Range (%) 87-102 81-92 85-97
      Average (%) 94 87 91

      Kit components & Storage

      The 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 since the kit is received.

      Item Specifications Storage
      Micro ELISA 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, 10 mL
      HRP Conjugate Diluent 1 vial, 10 mL
      Concentrated Wash Buffer (25×) 1 vial, 30 mL
      Substrate Reagent 1 vial, 10 mL 4℃(shading light)
      Stop Solution 1 vial, 10 mL 4℃
      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 in measuring instead of directly pouring.

      Other supplies required

      • Microplate reader with 450nm wavelength filter
      • 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.

      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 90 μL of Substrate Reagent. Incubate for 15 min at 37℃.

      6.Add 50 μL of Stop Solution.

      7.Read the OD at 450 nm immediately. Calculation of the results.

      Citations

      1. Publication: Zhao J, Xu S Z, Song F, et al. 2, 3, 5, 4' -Tetrahydroxystilbene-2-O-β-D-Glucoside Protects Human Umbilical Vein Endothelial Cells Against Lysophosphatidylcholine-Induced Apoptosis By Upregulating Superoxide Dismutase And Glutathione Peroxidase[J]. Iubmb Life, 2014, 66(10): 711-722.
        Species: Human
        Sample Type: cell lysis
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