Foot and Mouth Disease Virus Type AsiaⅠ Antibodies ELISA Kit

    • Animal Diseases ELISA Kit-Elabscience
    • Animal Diseases ELISA Kit-Elabscience
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    • Animal Diseases ELISA Kit-Elabscience
    • Animal Diseases ELISA Kit-Elabscience
    • Animal Diseases ELISA Kit-Elabscience
    • Animal Diseases ELISA Kit-Elabscience

      Catalog number:E-AD-E020

      Size:
      • 96T
      Qty:
      - +
      Price: $277

      Reactivity: Swine , Cattle, Goat, Sheep

      Sample type: Serum, plasma

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

      Add to cart Compare Bulk request Manual


      Test principle

      This kit is comprised by ELISA Microtiter plate pre-coated with FMDV-Asia-antibodies, HRP Conjugate and other auxiliary reagents, and apply the principle of Competitive-ELISA to detect FMDV-Asia-antibody in serum of sheep, cattle and goat. During the experiment, add Antigen Solution and samples into the ELISA Microtiter plate. Antibodies in sample will compete with antibody pre-coated on the Micro-plate for the antigen and block the combination between the antigen and the  Micro-plate. Then wash the plate to remove unbound antibodies and other components, add the Antibody Working Solution to incubate the reacting system. After washing, add the HRP Conjugate to specifically bind with the compound of antibody and antigen on the Micro-plate. The unbound HRP Conjugate will be removed by washing. Substrate is added into the well, it will react with the enzyme and the product become blue. The color shade is of negative correlation with specific antibody levels in the samples. At last, end the reaction by adding Stop Solution to produce a yellow product. Measure the absorbance value of each well by using a Micro-plate Reader with 450 nm wavelength, then we can know whether there are FMDV-Asia-antibodies in the samples.

       

      Kit components

      Item

      Specification

      ELISA Microtiter plate

      96 wells

      Dilution plate
      96 wells

      HRP Conjugate 

      5.5 mL

      Antibody Work Solution 

      5.5 mL

      Antigen Solution

      5.5 mL

      20×Concentrated Wash Buffer

      40 mL

      Substrate Reagent

      6 mL

      Substrate Reagent B

      6 mL

      Stop Solution

      6 mL

      Positive Control 

      1 mL

      Negative Control 

      1 mL

      Plate Sealer

      3 pieces

      Sealed Bag

      piece

      Manual

      1 copy

       

      Experimental instrument

      Microplate Reader with 450 nm wavelength filter or dual-wavelength (450/630 nm)

      High-precision transferpettor, EP tubes and disposable pipette tips

      37incubator or water bath

      Deionized or distilled water

      Absorbent paper


      Assay procedure
      1. Number: 

      Take out the Microplate, set 2 wells for negative/positive control respectively. The unused ELISA Microtiter plate should be sealed as soon as possible and stored at 2~8℃. Double well parallel experiment is recommended for detection.
      2. Add sample: 
       Add 50 μL of positive /negative control to positive/negative control wells. Add 25 µL of diluted wash buffer and add 25 µL of diluted sample to the sample wells. Then add 50 µL of Antigen Solution to each well. 

      Note: The dilution factor is 1:128 after adding antigen.

      3. Incubate: Gently tap the plate to ensure thorough mixing, incubate at 37℃ for 30 min.

      4. Wash: Aspirate each well and wash, repeat the process 5 times, immerse 30-60 sec each time. Wash by filling each well with wash buffer (approximately 300 μL). Complete removal of liquid at each step is essential. After the last wash, remove remained wash buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.

      5. Antibody Working Solution: Add 50 µL of Antibody Working Solution into each well (except the blank control well), and incubate at 37℃ for 30 min.

      6. Wash: Repeat Step 4 for washing.

      7. HRP conjugate: Add 100 µL of HRP conjugate into each well (except the blank control well), and incubate at 37℃ for 30 min.

      8. Wash: Repeat step 4 for washing.
      9. Color Development: Add 50 µL of substrate reagent A and 50 µL of substrate reagent B into each well, gently tap the plate to ensure thorough mixing, incubate for 15 min at 37℃. Protect from light.
      10. Stop reaction: Add 50 µL of stop solution into each well, gently mix. 、
      11. OD Measurement: Measure the absorbance value (A-value) of each well by using a Micro-plate reader with 450 nm wavelength (or use 630 nm as reference wavelength).



      Storage and Expiry date

      Store at 2~8with shading light for12 months.

      Please store the opened plate at 2~8, protect from light and moisture. The valid period is 2 months.

      Expiry date: expiration date is on the packing box.

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