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| Catalog # | Availability | Size | Quantity | Unit Price | Save For Later Wish List | |
|---|---|---|---|---|---|---|
| EKFT19315CN-48 | 2 weeks | 48T | $168.00 |
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| EKFT19315CN-96 | 2 weeks | 96T | $240.00 |
Product Overview | |
| Name | Canine Anti-JEV (Japanese Encephalitis Virus) IgM ELISA Kit |
|---|---|
| Sensitivity | Qualitative |
| Sample type(s) | Serum, Plasma |
| Principle of the Assay | |
| This kit was based on capture enzyme-linked immune-sorbent assay technology for the detection of anti-JEV IgM antibodies in Canine serum samples. Anti-Canine IgM（μ chain） was pre-coated onto 96-well plates. HRP-antigen was used as the detection conjugate. Controls and test samples were added to the appropriate wells, and then HRP-antigen was added. The HRP-antigen would bind to the anti-JEV IgM that had been captured by the Anti-Canine IgM（μ chain）. After washing away unbound components, TMB chromogenic substrate was added. The color development was read with a microplate reader, and the presence of anti-JEV IgM antibodies was determined according to the OD values. | |
| Sample Collection and Storage | |
| Isolate the test samples soon after collecting, then, analyze immediately (within 2 hours). Or aliquot and store at -20℃ for long term. Avoid multiple freeze-thaw cycles. Serum: Coagulate the serum at room temperature (about 1 hour). Centrifuge at approximately 1000 × g for 15 min. Analyze the serum immediately or aliquot and store at -20℃. Note: Samples to be used within 5 days may be stored at 2-8℃, otherwise samples must be stored at -20℃ (≤1 month) or -80℃(≤2 months) to avoid loss of bioactivity and contamination. Hemolyzed samples are not suitable for use in this assay. | |
| Material Required But Not Supplied | |
| 1. Microplate reader (wavelength: 450nm) 2.37℃ incubator 3. Automated plate washer 4. Precision single and multi-channel pipette and disposable tips 5. Clean tubes and Eppendorf tubes 6. Deionized or distilled water | |
| Assay procedure | |
| Remove the kit from the refrigerated environment and let it equilibrate at room temperature for 30 minutes before use 1. Label the sample wells, 3 Negative Controls, 2 Positive Controls and 1 blank well. 2. Add 50μL Negative Controls and Positive Controls to each well (except blank well). Add 50μL sample dilution buffer to sample wells and then add 5μL sample serum. 3. Add 50 μL of HRP-Conjugat to each well except the blank well. Gently tap the plate to ensure thorough mixing. Seal the plate with a cover and incubate at 37℃ for 30 min. 4. Remove the cover, and wash plate 5 times with Wash buffer and let the wash buffer stay in the wells for 0.5-1 minute each time. Wash the board 5 times in a row and pat dry the last time. 8. Add 90 μl of TMB substrate into each well. Gently tap the plate to ensure thorough mixing. Cover the plate and incubate at 37℃ in dark within 15 min. And the shades of blue can be seen in the Positive Controls. Negative Controls wells show no obvious color. 9. Add 50 μl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately. 10. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution. (Use the blank well to set zero)(Readings are taken within 30 minutes of termination of reaction) | |
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