Porcine Epidemic Diarrhea (PED) Antibody ELISA kit
1. Usage
This kit is used to detect Porcine Epidemic Diarrhea (PED) antibody in porcine serum.
2.Notes
1) All reagents should return to the room temperature(18~26℃) before using, and store back at 2-8℃after using
2) Do not use kit out of expiry date. Do not mix use reagents from kits of different lot numbers.
3) The unused micro-wells should seal back to bag and store back at 2-8℃.
4) Used materials should be treated innocuously, and be handled in accordance with local, regional and national regulations.
5) Avoid Substrate exposure to bright light, avoid contact with oxidants.
6) When the quantity of serum sample is big, dilute all serum sample on serum dilute plate firstly, then transfer all diluted serum sample on reaction wells, make the reaction time same.
7) Stop solution is corrosive, if splashed on the skin or clothing should immediately rinse with plenty of water.
8) Strictly adhere to instruction to get best result. All procedure including pipetting, timing and washing etc. must be accurate to get accurate result.
3.The kit components
| 1 | PED antigen coated micro-plate | 96T X 2 | |
| 2 | Enzyme conjugate | 22ml | yellow lid |
| 3 | Sample diluent solution | 50ml | transparent lid |
| 4 | PEDNegativecontrol serum | 1.5ml | green lid |
| 5 | PEDPositivecontrol serum | 1.5ml | red lid |
| 6 | Substrate | 12ml *2 | orange lid |
| 7 | Stop solution | 12ml | blue lid |
| 8 | 20×concentrated washing buffer | 50ml | white lid |
| 9 | Adhesive Foil | 2 pieces |
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| 10 | Instruction | 1 piece |
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4. Preparation
1) Sample dilute: Dilute sample with the sample diluent at 40 times.(5ul serum + 195ul sample diluent), the diluted sample need to mix evenly to get better results. Negative control and positive control do not need dilute.
2) Washing solution preparation: Dilute the 20×concentrated washing buffer with deionized water or distilled water at 20 times. (10ml 20×concentrated washing buffer + 190ml deionized water ) It is normal if there is crystallization in the 20×concentrated washing buffer, put at 37℃until completely dissolved.
5. ELISA procedure
1) Take pre-coated microtiter strips (Can unseal for several time use as per sample quantity), add 100μL diluted serum to test well, meanwhile set 2 wells for Negative control, 2 wells for Positive control separately. Add 100 μL Negative/Positive control to its well. Shake softly, cover andincubate at 37℃ for 30 min.
2) Pour the liquid out of the wells, add diluted washing solution to each well fully, be static for 10s, pour out. Wash3 times, at last pat to dry on absorbent paper.
3) Add 100 μL Enzyme Conjugate to each well, andincubate at 37℃ for 30 min.
4) Repeat the step 2(washing). Remember pat to dry on absorbent paper at last.
5) Add 100 μL substrate to each well, mix properly,react for 10 min at 37℃ at dark.
6) Add 50 μL stop solution in each well, and measure the result within 10 min(shake evenly before read result, no spill).
7) Read OD value with ELISA reader at 450nm/630nm.
6. Results
For the assay to be valid, the positive control wells’ average OD value must be greater than or equal to 0.6, and the negative control wells’ average OD value is less than 0.1. Otherwise the test is invalid, need test again.
The result is judged by S/P value,
S/P=(Sample OD450/630- NCx(—))/( PCx(—)- NCx(—)), NCx(—) means Negative control’s average OD450/630 value(calculate as 0.05 when the value is less than 0.05), PCx(—) means Positive control’s average OD450/630 value
If S/P≥0.2, it is positive; less than 0.2, it is negative.
Specifications: 96*2 wells/kit.
Expiry date:12 months.
Storage: Storing at 2-8℃, in the dark.