| Field | Specification |
|---|---|
| Applications | |
| Assay type | |
| Sample type(s) | Serum | Plasma |
| Detection method | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Product Overview
The Chicken IBV Antibody ELISA Kit detects antibodies against Chicken IBV in serum and plasma. The 96-well microplate is pre-coated, and the colorimetric signal is read at 450 nm.
Specifications
| Assay Type | Qualitative |
|---|---|
| Detection Method | Colorimetric (OD 450 nm) |
| Sample Type | Serum, Plasma |
| Kit Size | 96 T |
| Shipping | 2-8 °C |
Kit Components
| Component | Quantity |
|---|---|
| ELISA Microtiter plate | 96 wells |
| Dilution plate | 96 wells |
| HRP Conjugate | 11 mL |
| Sample Diluent | 50 mL |
| 20×Concentrated Wash Buffer | 40 mL |
| Substrate Reagent A | 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 | 1 piece |
| Manual | 1 copy |
Assay Procedure
Bring all reagents and samples to room temperature (25 °C) before use. All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. The unused ELISA Microtiter plate should be sealed as soon as possible and stored at 2-8 °C.
- Number the sample and control in order (in duplicate), and keep a record of control wells and sample wells. Set 2 wells for negative/positive control. Run samples in duplicate.
- Add 100 µL of positive/negative control to positive/negative control well. Add 10 µL of Diluted serum and 90 µL of Sample Diluent to the sample wells.
- Cover with a plate sealer and mix thoroughly, incubate at 37 °C for 30 min protected from light.
- Remove the liquid in each well. Immediately add 300 µL of Wash Buffer to each well and wash. Repeat 5 times, soaking 30 s each 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).
- Add 100 µL of HRP Conjugate to each well (except the blank control well), over the plate sealer and incubate at 37 °C for 30 min protected from light.
- Repeat step 4 for washing.
- Add 50 µL of Substrate Reagent A and 50 µL of Substrate Reagent B to each well. Cover with a plate sealer and mix thoroughly, incubate at 37 °C for 10 min protected from light.
- Add 50 µL of Stop Solution to each well, mix thoroughly.
- Measure the absorbance of each well by using a microplate reader with 450 nm (it is recommended to set the dual wavelength at 450 nm/630 nm) wavelength.
Required but not supplied: a microplate reader for 450 nm (dual wavelength 450 nm / 630 nm recommended), precision pipettes and tips, a 37 °C incubator or water bath, deionized or distilled water, and absorbent paper.
Result Interpretation
- Cut Off (C.O) =2.1× average of negative ODs (Take it as 0.1 if the average of negative ODs < 0.1; while average of negative ODs ≥ 0.1, calculate at the actual value).
- Positive result: ODs ≥ C.O
- Negative result: ODs < C.O
Storage & Handling
For Research Use Only. Not for use in diagnostic procedures.
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