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Avian Influenza Virus Antibodies ELISA Kit (AEES00734)

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SKU:
AEES00734
Product Type:
ELISA Kit
Size:
96 Assays
Target:
AIV-Ab
Reactivity:
Poultry
$719

Description

Product Name: Avian Influenza Virus Antibodies ELISA Kit
Target: AIV-Ab
SKU: AEES00734
Size: 96T
Reactivity: Poultry
Sample Type: Serum, Plasma, Yolk
Detection Method: Competitive ELISA
Assay Time: 75 min
Result Type: Qualitative; Sensitivity>98%, Specificity>98%
Detection Wavelength: 450/630nm
Storage: The majority of kits have a shelf-life of 12 months from the production date at 2-8℃.
Component Specification
ELISA Microtiter plate 96 Wells
HRP Conjugate 11 mL
25×Concentrated Wash Buffer 40 mL
Substrate Reagent A 6 mL
Substrate Reagent B 6 mL
Antibody Working Solution 6 mL
Stop Solution 6 mL
Positive Control 1 mL
Negative Control 1 mL
Plate Sealer 3 pieces
Plate Sealer 1 piece
Sealed Bag 1 copy

Other materials and equipment required:

  • Microplate Reader with 450nm wavelength filter or dual-wavelength (450/630nm)
  • High-precision transferpettor, EP tubes and disposable pipette tips
  • 37℃ incubator or water bath
  • Deionized or distilled water
  • Absorbent paper
  • Physiological saline solution

This kit is comprised by HRP conjugate, other auxiliary reagents, ELISA Microtiter plate pre-coated with the Avian Influenza Virus (AIV) antigen. Apply the principle of enzyme-linked immunoassay (ELISA) to detect AIV antibody in serum, plasma and yolk of poultry animals (chicken, duck, goose, etc.). During the experiment, add control serum and samples into the ELISA Microtiter plate. If AIV antibodies exist in the samples, it will compete with the antibody in the antibody working solution to bind with the antigen pre-coated on the Microplate. Then wash to remove unbound antibodies and other components, add the HRP conjugate to specifically bind with the compound of antibody and antigen on the microtiter plate. The unbound HRP conjugate will be removed by washing. Substrate Reagent is added into the well, it will react with the enzyme and become blue. The color shade is negative correlation with AIV 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 Microplate Reader with 450 nm wavelength, then we can judge whether AIV antibody exist in the sample.

Step 1: Number the sample and control in order (multiple well), and keep a record of control wells and sample wells. Set 2 wells for negative/positive control respectively. Samples need test in duplicate.
Step 2: Add sample: add 50 μL of positive/negative control to positive/negative control well, add 10 μL of Serum/plasma and 40 μL of Wash Buffer to each sample well.
Step 3: Incubate: add 50 μL of Antibody Working Solution to each well. Cover the plate sealer and mix thoroughly, incubate at 37℃ for 30 min in shading light.
Step 4: Wash: remove the liquid in each well. Immediately add 300 μL of Wash Buffer to each well and wash. Repeat wash procedure for 5 times, 30s intervals/time. Invert the plate and pat it against thick clean absorbent paper (If bubbles exist in the wells, clean tips can be used to prick them).
Step 5: HRP conjugate: add 100 µL of HRP Conjugate into each well, cover the plate sealer and incubate at 37℃ for 30 min in shading light.
Step 6: Wash: repeat step 4 for washing.
Step 7: Color Development: add 50 µL of Substrate Reagent A and 50 µL of Substrate Reagent B into each well, Cover the plate sealer and mix thoroughly, incubate at 37℃ for 15 min in shading light.
Step 8: Stop reaction: add 50 µL of Stop Solution into each well, mix thoroughly.
Step 9: OD Measurement: Measure the absorbance value (A-value) of each well by using a Microplate Reader with 450 nm wavelength (use 630 nm as reference wavelength).
Serum/plasma: Use the conventional method to prepare serum/plasma. The serum/plasma must be clear, no hemolysis, and no pollution. Samples can be conserved at 2-8 ℃ for 1 week, and it should be stored at -20℃ for long-term storage.
Yolk: Take 2 mL of fresh yolk and add 2 mL of physiological saline solution, oscillate to mix fully. Centrifuge at 3000 r/min for 15 min, take the supernatant for detection.
Wash Buffer: The 20× Concentrated Wash Buffer should be adjusted to room temperature to make the sediment dissolve fully before use, then dilute it with deionized water at 1:19.

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