Human ANP(Atrial Natriuretic Peptide) 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-H0532

      Synonyms:NPPA, ANF, ANP, ATFB6, ATRST2, CDD, CDD-ANF, CDP, PND, Atrial natriuretic peptide, natriuretic peptide A

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

      Reactivity: Human

      Detection Range: 7.81~500 pg/mL

      Sensitivity: 4.69 pg/mL

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

      Add to cart Compare Bulk request Manual

      Intended use

      This ELISA kit applies to the in vitro quantitative determination of Human ANP 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 Human ANP. During the reaction, Human ANP in the sample or standard competes with a fixed amount of Human ANP on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Human ANP. 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 Human ANP 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 Human
      Detection Method Colormetric
      Detection Range 7.81—500 pg/mL
      Sensitivity 4.69 pg/mL
      Sample Volume 50μL
      Sample Type Serum, plasma and other biological fluids

      Specificity

      This kit recognizes Human ANP in samples. No significant cross-reactivity or interference between Human ANP 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
      500 0.316
      0.348
      0.332
      250 0.406
      0.466
      0.436
      125 0.637
      0.615
      0.626
      62.5 0.924
      0.944
      0.934
      31.25 1.352
      1.352
      1.352
      15.63 1.809
      1.779
      1.794
      7.81 2.15
      2.166
      2.158
      0 2.677
      2.677
      2.677

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human ANP were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human ANP 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) 26.80 74.50 197.10 29.00 80.30 208.60
      Standard deviation 1.70 3.90 8.90 1.70 4.60 11.50
      C V (%) 6.34 5.23 4.52 5.86 5.73 5.51

      Recovery

      The recovery of Human ANP 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) 86-100 92
      EDTA plasma (n=5) 94-104 99
      Cell culture media (n=5) 92-107 99

      Linearity

      Samples were spiked with high concentrations of Human ANP 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 (%) 97-111 92-108 94-108
      Average (%) 103 99 99
      1:4 Range (%) 90-105 85-100 95-106
      Average (%) 96 92 100
      1:8 Range (%) 85-96 91-104 94-106
      Average (%) 90 97 100
      1:16 Range (%) 90-104 92-103 95-108
      Average (%) 96 97 101

      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

      1.Add 50 μL standard or sample to each well.

      2.Immediately add 50 μL Biotinylated Detection Ab to each well.

      3.Incubate for 45 min at 37℃. Aspirate and wash 3 times.

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

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

      6.Add 50 μL Stop Solution.

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

      Citations

      1. Publication: Soree P, Gupta R K, Singh K, et al. Raised Hif1Α During Normoxia In High Altitude Pulmonary Edema Susceptible Non-Mountaineers[J]. Scientific Reports, 2016.
        Sample Type: Plasma
      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.
        Sample Type: Serum
      3. Publication: Rani M R P, Anupama N, Sreelekshmi M, et al. Chlorogenic Acid Attenuates Glucotoxicity In H9c2 Cells Via Inhibition Of Glycation And PKC α Upregulation And Safeguarding Innate Antioxidant Status[J]. Biomedicine & Pharmacotherapy, 2018, 100: 467-477.
      4. Publication: Guzel D, Yazici A B, Pek T M, et al. atrial natriuretic peptide and posterior pituitary neurohormone changes in patients with acute schizophrenia[J]. Neuropsychiatric disease and treatment, 2018, 14: 1855.
        Sample Type: Human/Universal
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      • Q&A (1)
      Q

      M**********nSubmitted [ Jun 15 2018 ]

      Asked: Does this kit cross-react with mouse ANP?

      Reply

      A

      adminSubmitted [ Jun 15 2018 ]

      Answered: Hi Marina, Thanks for your inquiry. The cross-reactivity with the mouse ANP is 22.4% in our lab as the identity of the immunogen sequences is high. Any other questions, pls feel free to contact us. Best regards.

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