For scientific research purposes onlyIt shall not be used for medical diagnosis.

Carefully read this manual before use.

use way： Used for quantitative detection of pig serum, plasma, and related liquid samplesinterleukin2（IL-2）The content.

working principle

This kit uses a double site sandwich enzyme-linked immunosorbent assay (ELISA) method（ELISA）Determine the content in the samplePorcine interleukin2（IL-2）The level. Pre packagedPorcine interleukin2（IL-2）Add to the enzyme-linked well of the antibodyreference standardSamples to be tested andHRPmarkedinterleukin2（IL-2）Antibodies are incubated and washed to remove unbound components, and then substrates are addedATheB, producing blue,And it is converted into the final yellow color under the action of acid.The depth of color and the samplePorcine interleukin2（IL-2）concentrationPositive correlation.

Composition of reagent kit

1	Standard product（2000pg/ml）	0.5ml	7	chromogenic agentAliquid	6ml
2	Standard diluent	6mL	8	chromogenic agentBliquid	6ml
3	Enzyme labeled coated plate	12Kong×8item	9	Termination liquid	6ml
4	Enzyme linked immunosorbent assay (ELISA) reagents	6ml	10	instruction manual	1share
5	20Concentrated washing solution	25ml	11	sealing film	2Zhang
6	Sample diluent	6ml	12	sealed bag	1a

Note: The standard dilution solution for standard samples is sequentially diluted as follows:2000、1000、500、250、125The0 pg/ml

Reagents and equipment that are needed but not provided

1．37℃Thermostatic box.

2．Standard specification enzyme-linked immunosorbent assay (ELISA) reader.

3．Precision pipette and disposable suction head

4．Distilled water,

5．Disposable test tube

6．blotting paper

Precautions

1．from2-8The reagent kit taken out at ℃ should be equilibrated at room temperature at least before opening the kit30minute.If the enzyme label coated plate is not used up after opening, the Flat noodles shall be stored in a sealed bag.

2．Each step of sample addition should use a sampler and its accuracy should be regularly checked to avoid experimental errors

3．It is recommended that all standard samples and samples undergo double testing.

4．Strictly follow the instructions for operation, and the judgment of the test results must be based on the reading of the enzyme-linked immunosorbent assay reader.

5．To avoid cross contamination, it is important to avoid reusing the suction tips in your handssealing film.

6．Other unused reagents should be packaged or covered. Do not mix reagents of different batches. Use before shelf life.

7．substrateBSensitive to light, avoid prolonged exposure to light.

Washing method

Manual washing method: shake off the liquid inside the enzyme-linked immunosorbent assay plate; Lay several layers of absorbent paper on the experimental platform and pat the enzyme-linked immunosorbent assay (ELISA) plate downwards several times with force; Dilute the washing solution at least0.35mlInject into the hole and soak1-2minute. Repeat this process several times as needed.

Automatic board washing: If there is an automatic board washing machine, it should be used proficiently before being used in the formal experimental process

Specimen requirements

1．Cannot detect containingNaN3The sampleDue toNaN3Inhibition of horseradish peroxidase（HRP）Activity.

2．Extract the specimen as soon as possible after collection, according to relevant literature, and conduct experiments as soon as possible after extraction. If the experiment cannot be conducted immediatelyPlace the specimen in-20Store at ℃, but avoid repeated freeze-thaw cycles

operating procedure

1．Set up blank wells (blank control wells without sample or enzyme-linked immunosorbent assay, the rest of the steps are the same), standard wells, and test sample wells. Add standard to the standard well on the enzyme-linked immunosorbent assay (ELISA) coated plate50μlAdd sample diluent to the well of the sample to be tested on the enzyme-linked immunosorbent assay (ELISA) coated plate40μlThen add the sample to be tested10μlThe final dilution of the sample is5Double). Add enzyme labeled reagent to each well50μlExcept for blank holes. Gently shake and mix well,37℃ incubation60minute.

2．Discard the liquid, shake dry, fill each well with diluted washing solution, and shake30In seconds, shake off the detergent and pat dry with absorbent paper. So repeated5Next time, shoot dry.

3．Add color developer to each hole firstA50μlAdd color developer againB50μlGently shake and mix well,37Color development in the dark at ℃15minute.

4．Remove the enzyme-linked immunosorbent assay (ELISA) plate and add stop buffer to each well50μlTerminate the reaction (blue immediately turns yellow).

5．Measurement: Zero with blank hole, in450nmMeasure the absorbance values of each well under wavelength（ODValue). The measurement should be conducted after adding the termination solution15Conducted within minutes.

6．According to the concentration of the standard substance and its correspondingODCalculate the linear regression equation of the standard curve based on the value, and then use the sample'sODCalculate the corresponding sample concentration on the regression equation. Various application software can also be used for calculation. It should be remembered that due to the dilution of the sample, its actual concentration should be multiplied by the total dilution factor.

Summary of Operating Procedures：

Prepare reagents, samples, and standards

Add the prepared samples, standards, and enzyme-linked immunosorbent assay (ELISA) reagents,37℃ reaction60minute

Wash the board5Next, add the color developing solutionATheB，37Color development at ℃15minute

Add termination solution

15Read within minutesODvalue

calculate

Detection scope:31.2pg/ml-2000pg/ml.

Specifications 96T/box

preservation: 2-8℃

Validity period: 6A month（2-8℃）。

Shanghai Boyao Biotechnology Co., Ltdwww.afzhan.com/St37397