Pg(Progesterone) ELISA Kit

    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    <
    >
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience
    • competitive-ELISA-Elabscience

      Catalog number:E-EL-0154

      Size:
      • 96T
      • 24T
      Qty:
      - +
      Price: $495

      Reactivity: Universal

      Detection Range: 0.31~20 ng/mL

      Sensitivity: 0.15 ng/mL

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

      Add to cart Compare Bulk request

      Intended use

      This ELISA kit can be applied to the in vitro quantitative determination of Pg concentrations in serum, plasma, urine and saliva.

      Test principle

      This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with Pg. During the reaction, Pg in the sample or standard competes with a fixed amount of Pg on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Pg. Excess conjugate and unbound sample or standard are washed from the plate, and Avidin conjugated to Horseradish Peroxidase (HRP) are added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of Pg in the samples is then determined by comparing the OD of the samples to the standard curve.

      Assay type Competitive
      Format 96T
      Assay time 2.0h
      Reactivity Universal
      Detection Method Colormetric
      Detection Range 0.31—20 ng/mL
      Sensitivity 0.15 ng/mL
      Sample Volume 50μL
      Sample Type serum, plasma, urine and saliva

      Specificity

      This kit recognizes Pg in samples. No significant cross-reactivity or interference between Pg and analogues was observed.

      Typical data

      As the OD 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(ng/mL) O.D Average
      20 0.099
      0.083
      0.091
      10 0.153
      0.131
      0.142
      5 0.364
      0.266
      0.315
      2.5 0.415
      0.869
      0.642
      1.25 0.908
      0.926
      0.917
      0.62 1.451
      1.469
      1.46
      0.31 2.097
      2.191
      2.144
      0 2.995
      2.801
      2.898

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Pg were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Pg 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 (ng/mL) 0.56 2.22 8.78 0.54 2.01 8.55
      Standard deviation 0.05 0.14 0.53 0.05 0.14 0.53
      C V (%) 8.39 6.42 5.98 9.35 7.04 6.21

      Recovery

      The recovery of Pg 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) 80-94 84
      EDTA plasma (n=5) 82-96 87
      Urine (n=5) 85-109 95

      Linearity

      Samples were spiked with high concentrations of Pg 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) Urine (n=5)
      1:2 Range (%) 83-90 83-102 84-108
      Average (%) 85 90 93
      1:4 Range (%) 80-95 82-95 80-112
      Average (%) 89 89 92
      1:8 Range (%) 89-100 81-89 86-96
      Average (%) 92 86 87
      1:16 Range (%) 87-102 87-115 84-94
      Average (%) 91 98 88

      Kit components & Storage

      An unopened kit can be stored at 2-8°C 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 ELISA Plate(Dismountable) 8 wells ×12 strips -20°C, 6 months
      Reference Standard 2 vials
      Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 μL
      Concentrated HRP Conjugate (100×) 1 vial, 120 μL -20°C(shading light), 6 months
      Reference Standard & Sample Diluent 1 vial, 20 mL 2-8°C, 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 1 vial, 10 mL 2-8°C(shading light)
      Stop Solution 1 vial, 10 mL 2-8°C
      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

      • Microplate reader with 450 nm wavelength filter
      • High-precision transfer pipette, EP tubes and disposable pipette tips
      • Incubator capable of maintaining 37°C
      • Deionized or distilled water
      • Absorbent paper
      • Loading slot for Wash Buffer

      Assay Procedure

      elisa assay procedure 1

      1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C

      elisa assay procedure 2

      2. Aspirate and wash the plate for 3 times

      elisa assay procedure 3

      3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times

      elisa assay procedure 4

      4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C

      elisa assay procedure 5

      5. Add 50μL Stop Solution

      elisa assay procedure 6

      6. Read the plate at 450nm immediately. Calculation of the results

      Citations

      1. Publication: Yang X, Zhang W, Chen Y, et al. Activation Of Pparγ And Cd36 Expression--The Dual Pathophysiological Roles Of Progesterone[J]. Journal of Biological Chemistry, 2016.
        Species: Human
        Sample Type: Serum
      2. Publication: Zheng Q, Li Y L, Zhang D D, et al. ANP Promotes Proliferation And Inhibits Apoptosis Of Ovarian Granulosa Cells By NPRA/PGRMC1/EGFR Complex And Improves Ovary Functions Of PCOS Rats[J]. Cell Death & Disease, 2017.
        Species: Universal
        Sample Type: Serum
      3. Publication: Kong L H, Wang Q E, Jin J W, et al. Insulin Resistance Enhances The Mitogen-Activated Protein Kinase Signaling Pathway In Ovarian Granulosa Cells[J]. Plos One, 2017.
        Species: Universal
        Sample Type: Culture supernatant
      4. Publication: Yang C, Ye J, Liu Y, et al. Methylation pattern variation between goats and rats during the onset of puberty[J]. Reproduction in Domestic Animals, 2018.
        Species: Rat
      5. Publication: Wang M, Li L, Liu R, et al. Obesity-induced overexpression of miRNA-24 regulates cholesterol uptake and lipid metabolism by targeting SR-B1[J]. Gene, 2018, 668: 196-203.
        Species: Mouse
        Sample Type: Serum
      6. Publication: Song S , Tan Y . Expression of FKBP52 in the ovaries of PCOS rats[J]. International Journal of Molecular Medicine, 2018.
        Species: Rat
        Sample Type: Serum
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      Verified Customer

      Z*ySubmitted [ Jun 01 2019 ]

      • Species:Human
      • Sample:serum
      • Sample source:serum
      • Detection result:good
      • Description:This ELISA Kit was used to assay normal human(women and men) serum by us. We operated procedure as Elabscience,s the instruction manual provided and the experimental results were very good------when the human serums were detected without dilution, the testing results were in the range of standard curve, what’s more, It is very near compared with the clinical assay kit! The Pg level of normal men serum is 0.1-3.5 ng/mL and the follicular phase women serum is 0.1-2.1 ng/mL.
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