Swine encephalitis (JE) enzyme-linked immunoassay kit instructions - Database & Sql Blog Articles

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Swine Encephalitis (JE) Enzyme-Linked Immunosorbent Assay Kit

Instruction Manual

This kit is for research use only and not intended for human or animal diagnostic purposes.

Experimental Principle

The level of Japanese Encephalitis (JE) in the specimens is determined using the double antibody sandwich method. A microporous plate is coated with purified JE antibodies to create a solid-phase antibody. The JE antigen is then added to the microplate, followed by the addition of HRP-labeled JE antibodies. This forms an antibody-antigen-enzyme-labeled antibody complex. After thorough washing, the TMB substrate is added, resulting in a color change. Under the catalysis of HRP, TMB turns blue and then yellow when an acid is introduced. The intensity of the color is directly proportional to the concentration of JE in the sample. The absorbance (OD value) is measured at 450 nm using a microplate reader, and the JE concentration is calculated based on a standard curve.

Kit Composition

  1. 30× Washing Solution: 20ml × 1 bottle
  2. Stop Solution: 6ml × 1 bottle
  3. Enzyme Standard Reagent: 6ml × 1 bottle
  4. Standard (64pg/ml): 0.5ml × 1 bottle
  5. Enzyme-Labeled Coating Plate: 12 wells × 8 strips
  6. Standard Dilutions: 1.5ml × 1 bottle
  7. Sample Diluent: 6ml × 1 bottle
  8. Instructions: 1 copy
  9. Reagent A: 6ml × 1 bottle
  10. Reagent B: 6ml × 1 bottle
  11. Sealing Film: 2 sheets
  12. Sealed Bag: 1

Sample Requirements

1. Specimens should be processed as soon as possible after collection, following relevant protocols. If testing cannot be done immediately, store samples at -20°C, avoiding repeated freezing and thawing.

2. Samples containing NaN3 are not suitable for testing, as it may inhibit horseradish peroxidase (HRP) activity.

Testing Procedure

  1. Dilute the standard according to the provided instructions.
  2. Add 50μl of each standard and 50μl of the sample (after dilution) to the microplate.

Calculation

Plot the standard curve using standard concentrations and corresponding OD values. Use the linear regression equation to calculate the sample concentration from its OD value, and multiply by the dilution factor to obtain the actual concentration.

Precautions

  1. Allow the kit to reach room temperature before use. Store unopened enzyme-labeled reagents in a sealed bag.
  2. If the washing solution crystallizes, dissolve it in a water bath before use.
  3. Use a pipette for accuracy. Control loading time to avoid errors.
  4. Prepare a standard curve for each test, ideally in duplicate.
  5. Discard sealing film after single use to prevent contamination.
  6. Keep substrates away from light.
  7. Follow the manual strictly and rely on microplate reader results.
  8. Treat all samples and waste as biohazardous materials.
  9. Do not mix reagents from different batches.

Storage Conditions and Expiration Date

  1. Store the kit at 2–8°C.
  2. Shelf life: 6 months from the date of manufacture.

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