Avian EDS76 Antibody ELISA Test Kit for Poultry 96 Wells/Kit

1. Usage

This kit is used to detect Avian EDS76 virus antibody in chicken serum, to assess antibody condition by Avian EDS76 vaccine in chicken farm and assist diagnosis of serological infected chicken.

2. Principle

The Avian EDS76 antibody ELISA kit is based on an indirect enzyme immunoassay (indirect ELISA). Antigen is coated on the plate. When the sample serum contains specific antibodies against the virus, they bind to the antigen on the plate. Wash unbound antibody and other components. Specific enzyme conjugates are then added. After incubation and washing, TMB substrate is added. A colorimetric reaction will occur, measured by a spectrophotometer (450 nm).

3. Reagents

4. Materials required but not provided

1) Micropipettors and disposable tips: 0.5μL~10μL,10μL~100μL,100μL~1000μL
2) Disposable tips
3) 37 ℃ Incubator
4) Measuring cylinder: 500 ml
5) 96 wells microplate reader
6) Distilled water or deionied water
7) Bottle or microplate washing machine

5. Sample preparation

Whole blood is taken from animals, and serum is prepared according to conventional methods. The serum should be clear without hemolysis.

6. Preparation of washing buffer

Return the wash buffer to room temperature before use. If there are salty crystals, shake to dissolve the crystals, and then dilute 10-fold with distilled or deionized water. The diluted wash buffer can be stored at 4°C for 1 week.

7. Sample dilution

At serum dilution plate, dilute serum at 1:99 with sample dilution (for example: 495μL sample dilution + 5μL serum)
Notice: Negative control serum and Positive control serum do not need dilute. Exchange tip after taking sample every time, record the situation of the sample on plate accurately. Shake the sample evenly before adding it.

8. Notes

1) All reagents should be adjusted to the room temperature and shake evenly before using, store at 2-8 ℃after using
2) Do not exchange the reagents from the kits of different lot numbers to use. Avoid reagent pollution when using.
3) Substrate and stop solution may be excitant to skin and eyes, pay attention when using.
4) Do not expose TMB (Substrate B) to light and avoid it contact with antioxidants.
5) The wells should avoid damp or touching water after unsealing (Put the un-using microplate back to bag with dehydrator in 2~8 ℃soon )
6) Deal all waste reasonable before dumping to avoid pollution.
7) Strictly adhere to instruction to get best result. All procedure including pipetting, timing and washing etc. must be accurate.
8) Serum dilution plate is disposable, do not use for second time; the MAX volume of it is 300μL/well.

9. ELISA procedure

1) Take pre-coated microplate (Can unseal for several time use as per sample quantity), add 100μL diluted serum to a well, meanwhile set 1 wells for Negative control serum, Positive control serum and blank control wells separately. Add 100 μL Negative/Positive control serum to its wells, only add 100μL sample dilution in the blank control well. Shake softly, do not overflow,incubate at 37℃ for 30 min.
2) Pour the liquid out of the wells, add 250 μL diluted washing solution to each well, static for 1 min, pour out. Repeat 3 times, then pat to dry on absorbent paper.
3) Add 100μL Enzyme conjugate to each well, shake softly andincubate at 37℃ for 30 min.
4) Repeat the step 2(washing). Remember pat to dry on absorbent paper at last.
5) Add 50 μL substrate A, then substrate B (50 μL) to each well, mix properly,react for 10 min atdark at37℃ in dark.
6) Add 50 μl of stop solution in each well, and measure the result within 10 min.

10. Results

Set to zero at blank control wells and test the OD450nm (630 nm as reference) value on a microplate reader. The conditions for the establishment of the test are that the average OD450nm value of the positive control wells is greater than or equal to 0.6, and the average OD450nm value of the negative control wells is less than 0.15. If the test is invalid and the operating procedure is skeptical, re-run the test and observe all reagents carefully.
If the A450 value of the sample is greater than 0.2+ absorbance of the negative control, it is judged as positive; if the absorbance of the negative control is less than 0.2+, it is negative. If the absorbance of the negative control is less than 0.05, calculate as 0.05

Specifications: 96 wells/kit.
Expiry date:12 months.
Storage: Storing at 2-8℃, in the dark.

Q1: How long will it take for you to ship?
A1: Usually ships within 7 working days.

Q2: Do you support OEM/ODM?
A2: can be supported. We can customize according to your specific needs and specific quantities.

Q3: How is your factory doing in terms of quality control?
A3: We have ISO9001 certification and ISO13485 certification, so far all the production process, we have standard rules, we comply with the relevant behavior and laws of the government.

Q4: Is after-sales service guaranteed?
A4: We provide professional online technical after-sales service. If the product fails during the experiment, we can provide
one-to-one guidance through telephone, video and other forms.

Q5: What is your minimum order quantity?
A5: 1Kit.

Q6: What is the shipping method?
A6: It can be shipped by express (FEDEX, UPS, DHL, EMS, etc.) or by air and ground.Please confirm with us before placing an order.