NA/NE(Noradrenaline/Norepinephrine) 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-0047

      Synonyms:Norepinephrine, NE, noradrenaline, NA, NAd, norad, 4, 5-β-trihydroxyphenethylamine

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

      Reactivity: Universal

      Detection Range: 0.31~20 ng/mL

      Sensitivity: 0.19 ng/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  NA/NE 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 NA/NE. During the reaction,  NA/NE in the sample or standard competes with a fixed amount of NA/NE on the solid phase supporter for sites on the Biotinylated Detection Ab specific to  NA/NE. 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  NA/NE in the samples is then determined by comparing the OD of the samples to the standard curve.

      Assay type Competitive-ELISA
      Format 96T
      Assay time 2.5h
      Reactivity Universal
      Detection Method Colormetric
      Detection Range 0.31—20 ng/mL
      Sensitivity 0.19 ng/mL
      Sample Volume 50μL
      Sample Type Serum, plasma and other biological fluids

      Specificity

      This kit recognizes NA/NE in samples. No significant cross-reactivity or interference between NA/NE 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 0.32
      0.342
      0.331
      10 0.444
      0.45
      0.447
      5 0.654
      0.648
      0.651
      2.5 0.956
      0.958
      0.957
      1.25 1.339
      1.309
      1.324
      0.63 1.66
      1.658
      1.659
      0.31 1.9
      1.914
      1.907
      0 2.196
      2.21
      2.203

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level NA/NE were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level NA/NE 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) 1.10 2.30 9.50 1.00 2.40 9.60
      Standard deviation 0.10 0.10 0.30 0.10 0.10 0.40
      C V (%) 9.09 4.35 3.16 10.00 4.17 4.17

      Recovery

      The recovery of  NA/NE 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) 88-99 93
      EDTA plasma (n=5) 94-110 102
      Cell culture media (n=5) 89-102 97

      Linearity

      Samples were spiked with high concentrations of NA/NE 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 (%) 89-105 99-112 99-114
      Average (%) 96 105 106
      1:4 Range (%) 87-102 90-101 95-111
      Average (%) 94 96 102
      1:8 Range (%) 86-99 90-103 96-110
      Average (%) 92 97 102
      1:16 Range (%) 92-107 86-98 97-112
      Average (%) 97 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

      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: Li D, Wang Q, Zhang Y, et al. A Novel Swine Model Of Spontaneous Hypertension With Sympathetic Hyperactivity Responds Well To Renal Denervation[J]. American Journal of Hypertension, 2016, 29(1): 63-72.
        Sample Type: Plasma and urine
      2. Publication: Lu W, Kang J, Hu K, et al. The Role Of The Nox4-Derived ROS-Mediated RhoA/Rho Kinase Pathway In Rat Hypertension Induced By Chronic Intermittent Hypoxia[J]. Sleep and Breathing, 2017.
        Sample Type: Serum and tissue homogenates
      3. Publication: He J L, Zhao M, Xia J J, et al. FGF21 Ameliorates The Neurocontrol Of Blood Pressure In The High Fructose-Drinking Rats[J]. Scientific Reports, 2016.
        Sample Type: Serum
      4. Publication: Wang K, Lu D, Zhang B, et al. Renal Denervation Attenuates Multi-Organ Fibrosis And Improves Vascular Remodeling In Rats With Transverse Aortic Constriction Induced Cardiomyopathy.[J]. Cellular Physiology and Biochemistry, 2016, 40: 465-476.
        Sample Type: Plasma and tissue homogenates
      5. Publication: Yu L L, Li X F, Huang B, et al. Atrial Fibrillation In Acute Obstructive Sleep Apnea: Autonomic Nervous Mechanism And Modulation[J]. Journal of the American Heart Association, 2017.
        Sample Type: Plasma
      6. Publication: Lü Y F, Yang Y, Li C L, et al. The Locus Coeruleus-Norepinephrine System Mediates Empathy For Pain Through Selective Up-Regulation Of P2X3 Receptor In Dorsal Root Ganglia In Rats[J]. Frontiers in Neural Circuits, 2017.
        Sample Type: Serum
      7. Publication: Tang C L, Huang X, Kang F, et al. Intranasal Dexmedetomidine On Stress Hormones, Inflammatory Markers, And Postoperative Analgesia After Functional Endoscopic Sinus Surgery[J]. Mediators Of Inflammation, 2015.
        Sample Type: Serum
      8. Publication: Yu X, He W, Qin Z, et al. Selective ablation of the ligament of Marshall attenuates atrial electrical remodeling in a short‐term rapid atrial pacing canine model[J]. Journal of cardiovascular electrophysiology, 2018.
        Sample Type: Dog
      9. Publication: Zhang B, Li X, Chen C, et al. Renal Denervation Effects on Myocardial Fibrosis and Ventricular Arrhythmias in Rats with Ischemic Cardiomyopathy.[J]. Cellular Physiology and Biochemistry, 2018, 46(6):2471-2479.
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