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    Home > Biochemistry News > Biotechnology News > Operation steps of monkey pox virus antibody (PV-Ab) ELISA kit

    Operation steps of monkey pox virus antibody (PV-Ab) ELISA kit

    • Last Update: 2022-08-30
    • Source: Internet
    • Author: User
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    Monkeypox virus antibody (PV-Ab) ELISA kit operating steps​Detection principle Sample collection, processing and storage methods Monkeypox virus antibody (PV-Ab) ELISA kit operating steps​Self-prepared items
    • Microplate reader (450nm)
    • High-precision sampler and pipette tip: 0.
      5-10uL, 2-20uL, 20-200uL, 200-1000uL
    • 37°C incubator

    Operation Precautions
    1. The kit was stored at 2-8°C and equilibrated at room temperature for 20 minutes before use
      .
      There will be crystals in the concentrated washing liquid taken out from the refrigerator, which is a normal phenomenon, and the crystals are completely dissolved by heating in a water bath before use
      .
    2. The slats not used in the experiment should be immediately put back into the ziplock bag and stored in an airtight (low temperature drying)
      .
    3. The pretreated sample should be properly diluted with sample diluent according to the operation steps to achieve the best detection effect of the kit
      .
    4. Incubate in strict accordance with the time, volume and sequence indicated in the instructions
      .
    5. Shake all liquid components well before use
      .

    Kit composition
    name 96-well configuration 48 well configuration Remark
    microtiter plate 12 holes × 8 strips 12 holes × 4 strips none
    negative control 0.
    5mL
    0.
    5mL
    none
    positive control 0.
    5mL
    0.
    5mL
    none
    Detection of antigen-HRP 10mL 5mL none
    20x wash buffer 25mL 15mL Dilute according to instructions
    Sample Diluent 6mL 3mL none
    Substrate A 6mL 3mL none
    Substrate B 6mL 3mL none
    Stop solution 6mL 3mL none
    Sealing film 2 sheets 2 sheets none
    manual 1 serving 1 serving none
    Ziplock bag 1 1 none

    Monkeypox virus antibody (PV-Ab) ELISA kit operation steps​Reagent preparation and washing method
    1. Manual plate washing: shake off the liquid in the wells, fill each well with washing liquid, let stand for 1 min, shake off the liquid in the wells, pat dry on absorbent paper, and wash the plate 5 times
      .
    2. Automatic plate washer: inject 350 μL of washing solution into each well, soak for 1 min, and wash the plate 5 times
      .

    Steps
    1. Take out the required slats from the aluminum foil bag after equilibrating at room temperature for 20 min, and seal the remaining slats in a ziplock bag and put them back at 4°C
      .
    2. Set negative and positive control wells and sample wells, and add 50 μL of negative control and positive control to each of the negative and positive control wells;
    3. First add 10 μL of the sample to be tested, and then add 40 μL of the sample diluent to the well of the sample to be tested;
    4. Subsequently, 100 μL of horseradish peroxidase (HRP)-labeled detection antigen was added to each well of the negative and positive control wells and sample wells, and the reaction wells were sealed with a sealing film, and incubated at 37°C in a water bath or incubator for 60 min
      .
    5. Discard the liquid, pat dry on absorbent paper, fill each well with washing solution, let stand for 1 min, shake off the washing solution, pat dry on absorbent paper, and repeat the plate washing 5 times (you can also use a plate washer to wash the plate)
      .
    6. Add 50 μL each of substrate A and B to each well, and incubate at 37°C for 15 min in the dark
      .
    7. Add 50 μL of stop solution to each well, and measure the OD value of each well at a wavelength of 450 nm within 15 min
      .

    The results judge the performance of the kit
    •  Accuracy: The average OD value of the positive control wells is ≥1.
      00; the average OD value of the negative control wells is ≤0.
      15, indicating that the test results are valid
      .
    •  Specificity: Does not cross-react with other soluble structural analogs
      .
    •  Repeatability: The coefficient of variation within and between plates is less than 15%
      .
    •  Storage: 2-8℃, keep away from light and moisture
      .
    •  Validity: 6 months

    Disclaimer
    •   The kit is for research use only, and shall not be used in clinical experiments or human experiments.
      Otherwise, the experimenter shall bear all the consequences, and the company shall not be responsible
      .
    •   Strictly follow the instructions, if the experimenter violates the instructions, the consequences will be borne by the experimenter
      .
      Monkeypox virus antibody (PV-Ab) ELISA kit operation steps​
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