| Field | Specification |
|---|---|
| Applications | |
| Assay type | |
| Sample type(s) | Serum | Plasma |
| Sensitivity | |
| Detection range | |
| Detection method | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Product Overview
The Porcine JEV Antibody ELISA Kit is an indirect ELISA kit for quantitative measurement of porcine antibodies against JEV in serum and plasma. The 96-well microplate is pre-coated, and the colorimetric signal is read at 450 nm.
In this indirect format, porcine antibodies against JEV in the sample bind in the pre-coated wells and are then detected with an enzyme-labeled reagent, so the optical density rises with the antibody level. Results are calculated from the standard curve run on the same plate.
Values are expressed relative to the antibody standard supplied with the kit.
Specifications
| Assay Type | Indirect (Quantitative) |
|---|---|
| Detection Method | Colorimetric (OD 450 nm) |
| Sample Type | Serum, Plasma |
| Sensitivity | 0.25 ng/mL |
| Detection Range | 1.56 - 100 ng/mL |
| Recovery | 80-120% |
| Kit Size | 96 T |
| Shipping | 2-8 °C |
Kit Components
| Component | Quantity |
|---|---|
| ELISA Microtiter plate | 96 wells*2 |
| HRP Conjugate | 24 mL |
| Sample Diluent | 100 mL |
| 10×Concentrated Wash Buffer | 100 mL |
| Substrate Reagent A | 12 mL |
| Substrate Reagent B | 12 mL |
| Stop Solution | 12 mL |
| Positive Control | 1.5 mL |
| Negative Control | 1.5 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. Set 1 blank control well， Run samples in duplicate.
- Add 100 µL of positive/negative control to positive/negative control well, add 100 µL of Sample Diluent to the blank well, and add 100 µL of diluted serum 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 350 µL of Wash Buffer to each well and wash. Repeat 3 times, soaking 60 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, cover with a 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. (Add Substrate Reagent A first and then Substrate Reagent B, please add in order) 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.
Storage & Handling
For Research Use Only. Not for use in diagnostic procedures.
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