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Avian Influenza Virus H9 Antibodies ELISA Kit (AEES00736)

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SKU:
AEES00736
Product Type:
ELISA Kit
Size:
96 Assays
Target:
AIV-H9-Ab
Reactivity:
Poultry
€599

Description

Product Name: Avian Influenza Virus H9 Antibodies ELISA Kit
Target: AIV-H9-Ab
SKU: AEES00736
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 (0.9%)

This kit is comprised by HRP conjugate, other reagents, ELISA Microtiter plate pre-coated with the Avian Influenza Virus H9 (AIV-H9) antigen. Apply the principle of enzyme-linked immunoassay (ELISA) to detect AIV-H9-Ab in serum, plasma and yolk of poultry animals. During the experiment, add control, samples, and antibody working solution into the ELISA microtiter plate, AIV-H9-Ab will compete with the antibody working solution to bind with the antigen pre-coated on the ELISA Microtiter plate. Then horseradish peroxidase (HRP) conjugate is added to each ELISA microtiter plate well, and substrate reagent is added for color development. There is a negative correlation between the OD value of samples and the concentration of AIV-H9-Ab. Measure the absorbance value of each well by using a microplate reader with 450 nm (630 nm) wavelength, then we can judge whether AIV-H9 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 sample 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, 30 s 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 or plasma, the serum or plasma must be clear, no hemolysis and no pollution. Samples can be conserved at 2-8℃ in 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 analysis.
Wash Buffer: The 20× Concentrated Wash Buffer should be adjusted to room temperature before use, then dilute it with deionized or distilled water at 1:19.

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