Rat ET-1(Endothelin 1) 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-R0167

      Synonyms:EDN1, ET1, HDLCQ7, PPET1, Preproendothelin-1

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

      Reactivity: Rat

      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 Rat ET-1 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 Rat ET-1. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat ET-1 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 Rat ET-1, 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 Rat ET-1. You can calculate the concentration of Rat ET-1 in the samples by comparing the OD of the samples to the standard curve.

      Assay type Sandwich
      Format 96T
      Assay time 4.5h
      Reactivity Rat
      Detection Method Colormetric
      Detection Range 7.81—500 pg/mL
      Sensitivity 4.69 pg/mL
      Sample Volume Required Per Well 100μL
      Sample Type Serum, plasma and other biological fluids

      Specificity

      This kit recognizes Rat ET-1 in samples. No significant cross-reactivity or interference between Rat ET-1 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
      500 2.34
      2.386
      2.363 2.296
      250 1.686
      1.712
      1.699 1.632
      125 0.927
      0.923
      0.925 0.858
      62.5 0.478
      0.492
      0.485 0.418
      31.25 0.261
      0.261
      0.261 0.194
      15.63 0.174
      0.164
      0.169 0.102
      7.81 0.116
      0.122
      0.119 0.052
      0 0.062
      0.072
      0.067 --

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat ET-1 were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat ET-1 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) 24.70 70.80 167.70 22.60 75.30 173.10
      Standard deviation 1.30 2.90 6.40 1.30 3.10 7.10
      C V (%) 5.26 4.10 3.82 5.75 4.12 4.10

      Recovery

      The recovery of Rat ET-1 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) 85-96 90
      EDTA plasma (n=5) 88-99 93
      Cell culture media (n=5) 88-102 93

      Linearity

      Samples were spiked with high concentrations of Rat ET-1 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 (%) 96-111 99-111 89-102
      Average (%) 102 105 94
      1:4 Range (%) 96-111 87-100 92-107
      Average (%) 104 94 98
      1:8 Range (%) 101-115 84-99 90-106
      Average (%) 108 91 97
      1:16 Range (%) 101-116 81-94 92-106
      Average (%) 107 86 99

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

      2.Remove the liquid.

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

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

      5.Add 90 μL of 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: Kurt A, Tumkaya L, Kalkan Y, et al. Is Adalimumab Protective In Ischemia-Reperfusion Injury In Lung[J]. Iranian Journal of Basic Medical Sciences, 2015, 18(11): 1093-1099.
        Sample Type: Tissue homogenates
      2. Publication: Zhang K, Wang D, Jiang Z, et al. Endothelial Dysfunction Correlates With Decompression Bubbles In Rats[J]. Scientific Reports, 2016.
        Sample Type: Serum
      3. Publication: Damiani E, Ince C, Orlando F, et al. Effects Of The Infusion Of 4% Or 20% Human Serum Albumin On The Skeletal Muscle Microcirculation In Endotoxemic Rats[J]. Plos One, 2016.
        Sample Type: Serum
      4. Publication: Li G, Peng Y, Zhao T, et al. Plumbagin Alleviates Capillarization Of Hepatic Sinusoids In Vitro By Downregulating ET-1, VEGF, LN, And Type IV Collagen[J]. BioMed Research International, 2017.
        Sample Type: culture supernatant
      5. Publication: Kurt A, Tumkaya L, Turut H, et al. Protective Effects Of Infliximab On Lung Injury Induced By Methotrexate[J]. Archivos De Bronconeumología (English Edition), 2015, 51(11): 551-557.
        Sample Type: Tissue homogenate
      6. Publication: 81-393 Gdynia Abrahama 68/4 Poland
        Sample Type: Tissue Homogenates
      7. Publication: Bellaye P S, Yanagihara T, Granton E, et al. Macitentan reduces progression of TGF-β1-induced pulmonary fibrosis and pulmonary hypertension[J]. European Respiratory Journal, 2018: 1701857.
        Sample Type: Rat
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