Rat DPD(Deoxypyridinoline) ELISA Kit

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

      Catalog number:E-EL-R0327

      Synonyms:Pyrilinks-D, deoxyPYD

      Size:
      • 96T
      • 24T
      Qty:
      - +
      Price: $609

      Reactivity: Rat

      Detection Range: 3.13~200 ng/mL

      Sensitivity: 1.88 ng/mL

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

      Add to cart Compare Bulk request

      Intended use

      This ELISA kit applies to the in vitro quantitative determination of Rat DPD concentrations in serum, plasma and other biological fluids.

      Test principle

      This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with Rat DPD. During the reaction, Rat DPD in the sample or standard competes with a fixed amount of Rat DPD on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Rat DPD. 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 Rat DPD 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 Rat
      Detection Method Colormetric
      Detection Range 3.13—200 ng/mL
      Sensitivity 1.88 ng/mL
      Sample Volume 50μL
      Sample Type Serum, plasma and other biological fluids

      Specificity

      This kit recognizes Rat DPD in samples. No significant cross-reactivity or interference between Rat DPD 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
      200 0.375
      0.397
      0.386
      100 0.507
      0.523
      0.515
      50 0.74
      0.72
      0.73
      25 1.04
      1.042
      1.041
      12.5 1.421
      1.403
      1.412
      6.25 1.774
      1.752
      1.763
      3.13 2.025
      2.033
      2.029
      0 2.405
      2.409
      2.407

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat DPD were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat DPD 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) 9.70 28.00 83.70 9.80 29.70 77.20
      Standard deviation 0.60 1.70 2.70 0.60 1.80 3.50
      C V (%) 6.19 6.07 3.23 6.12 6.06 4.53

      Recovery

      The recovery of Rat DPD 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) 90-102 95
      EDTA plasma (n=5) 86-97 91
      Cell culture media (n=5) 95-108 101

      Linearity

      Samples were spiked with high concentrations of Rat DPD 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 (%) 94-106 89-102 90-104
      Average (%) 99 96 96
      1:4 Range (%) 84-94 93-105 93-106
      Average (%) 89 98 100
      1:8 Range (%) 83-97 85-97 95-108
      Average (%) 89 92 100
      1:16 Range (%) 86-99 92-106 95-107
      Average (%) 92 98 101

      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: Deng Y, Kang W, Zhao J, et al. Osteoprotective Effect Of Echinocystic Acid, A Triterpone Component From Eclipta Prostrata, In Ovariectomy-Induced Osteoporotic Rats[J]. Plos One, 2015.
        Species: Rat
        Sample Type: Serum
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