Human PⅢNP(N-Terminal Procollagen Ⅲ Propeptide) ELISA Kit

    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • 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-H0183

      Synonyms:P3NP, N-Propeptide Of Type Ⅲ Procollagen, Procollagen Ⅲ Amino Terminal Propeptide

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

      Reactivity: Human

      Detection Range: 23.44~1500 pg/mL

      Sensitivity: 14.06 pg/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 Human PⅢNP concentrations in serum, plasma and other biological fluids.

      Test principle

      This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human PⅢNP. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human PⅢNP 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 Human PⅢNP, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of 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 PⅢNP. You can calculate the concentration of Human PⅢNP in the samples by comparing the OD of the samples to the standard curve.

      Assay type Sandwich
      Format 96T
      Assay time 3.5h
      Reactivity Human
      Detection Method Colormetric
      Detection Range 23.44—1500 pg/mL
      Sensitivity 14.06 pg/mL
      Sample Volume Required Per Well 100μL
      Sample Type Serum, plasma and other biological fluids

      Specificity

      This kit recognizes Human PⅢNP in samples. No significant cross-reactivity or interference between Human PⅢNP 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
      (pg/mL)
      O.D Average Corrected
      1500 2.438
      2.464
      2.451 2.381
      750 1.724
      1.764
      1.744 1.674
      375 0.947
      0.925
      0.936 0.866
      187.5 0.517
      0.553
      0.535 0.465
      93.75 0.289
      0.285
      0.287 0.217
      46.88 0.188
      0.16
      0.174 0.104
      23.44 0.12
      0.126
      0.123 0.053
      0 0.061
      0.079
      0.07 --

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human PⅢNP were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human PⅢNP 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) 75.40 208.10 518.30 69.80 205.90 509.50
      Standard deviation 4.80 11.40 18.70 3.90 9.90 18.30
      C V (%) 6.37 5.48 3.61 5.59 4.81 3.59

      Recovery

      The recovery of Human PⅢNP 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) 95-107 101
      EDTA plasma (n=5) 96-107 102
      Cell culture media (n=5) 85-99 92

      Linearity

      Samples were spiked with high concentrations of Human PⅢNP 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 (%) 94-109 86-98 90-103
      Average (%) 101 93 97
      1:4 Range (%) 96-106 87-102 95-110
      Average (%) 101 93 101
      1:8 Range (%) 99-113 83-96 92-108
      Average (%) 105 89 99
      1:16 Range (%) 94-106 88-101 98-111
      Average (%) 100 93 104

      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 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, 14 mL
      HRP Conjugate Diluent 1 vial, 14 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 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℃
      • Deionized or distilled water
      • Absorbent paper
      • Loading slot for Wash Buffer

      Assay Procedure

      elisa assay procedure 1

      1.Add 100 μL standard or sample to each well. Incubate for 90 min at 37℃.

      elisa assay procedure 2

      2.Remove the liquid.

      elisa assay procedure 3

      3.Add 100 μL Biotinylated Detection Ab. Incubate for 1 hour at 37℃. Aspirate and wash 3 times.

      elisa assay procedure 4

      4.Add 100 μL HRP Conjugate. Incubate for 30 min at 37℃. Aspirate and wash 5 times.

      elisa assay procedure 5

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

      elisa assay procedure 6

      6.Add 50 μL Stop Solution.

      elisa assay procedure 7

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

      Citations

      1. Publication: Liu R, Chen L, Wei W, et al. Extracellular Matrix Turnover In Coronary Artery Ectasia Patients[J]. Heart and Vessels, 2015.
        Sample Type: Serum
      2. Publication: Begg G A, Lip G Y, Plein S, et al. Circulating Biomarkers Of Fibrosis And Cardioversion Of Atrial Fibrillation: A Prospective, Controlled Cohort Study[J]. Clinical Biochemistry, 2016.
        Sample Type: Serum
      3. Publication: Barchetta I, Del B M, Angelico F, et al. No Effects Of Oral Vitamin D Supplementation On Non-Alcoholic Fatty Liver Disease In Patients With Type 2 Diabetes: A Randomized, Double-Blind, Placebo-Controlled Trial[J]. BMC Medicine, 2016.
        Sample Type: Serum
      4. Publication: Wu J, Wang K, Xu J, et al. Associations Between Serum Ghrelin And Knee Symptoms, Joint Structures And Cartilage Or Bone Biomarkers In Patients With Knee Osteoarthritis[J]. Osteoarthritis and Cartilage, 2017.
        Sample Type: Serum
      5. Publication: Barchetta I, Cimini F A, De Gioannis R, et al. Circulating PIIINP Levels Identify Adipose Tissue‐Associated Inflammation In Individuals With Type 2 Diabetes Mellitus With And Without Non‐Alcoholic Fatty Liver Disease[J]. Diabetes/Metabolism Research And Reviews, 2018.
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