This kit is for research use only.

Detection range:96T

2pg/ml-60pg/ml

Purpose of use:

This reagent kit is used for the determination of serum, plasma, and related liquid samples in miceBinterferon(IFN-(b)Content.

Experimental Principle

This test kit uses the double antibody sandwich method for determinationspecimenmiddlemouseBinterferon(IFN-(b)level Using purifiedmouseBinterferon(IFN-(b)Antibody coated microplates are used to prepare solid-phase antibodies, which are then sequentially added to the micropores coated with monoclonal antibodiesBinterferon(IFN-(b)And then withHRPmarkedBinterferon(IFN-(b)Antibody binding, forming antibodies-antigen-Enzyme linked antibody complexAfter washingplussubstrateTMBColor rendering.TMBinHRPUnder the catalysis of enzymes, it is converted into blue, and under the action of acid, it is converted into the final yellow. The depth of color and the sampleBinterferon(IFN-(b)Positive correlation. Using an enzyme-linked immunosorbent assay (ELISA) reader450nmMeasure absorbance at wavelength（ODValue),By standard curveCalculate the sampleSmall and medium-sized miceBinterferon(IFN-(b)Concentration.

Kit components

1	30Concentrated washing solution	20ml×1bottle	7	Stop Solution	6ml×1bottle
2	Enzyme linked immunosorbent assay (ELISA) reagents	6ml×1bottle	8	standardProduct（120pg/ml）	0.5ml×1bottle
3	enzymeThe package is labeled asboard	12Kong×8item	9	reference standarddiluent	1.5ml×1bottle
4	samplediluent	6ml×1bottle	10	Instruction manual	1copy
5	Chromogenic agentAliquid	6ml×1bottle	11	Sealing film	2Zhang
6	Chromogenic agentBliquid	6ml×1/bottle	12	sealed bag	1a

Specimen requirements

1Extract 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 immediately, the specimen can be placed in-20Stored at ℃, but shouldAvoid repeated freeze-thaw cycles

2. Samples containing NaN3 cannot be detected becauseNaN3Inhibition of horseradish peroxidase（HRP）Activity.

operating steps

Dilution of standard samples: This kit provides one original standard sample, and users can dilute it in a small test tube according to the following chart.

60pg/ml	5Standard product number	150MlAddition of original standard samples150MlStandard diluent
30pg/ml	4Standard product number	150Mlof5Number of standard products added150MlStandard diluent
15pg/ml	3Standard product number	150Mlof4Number of standard products added150MlStandard diluent
7.5pg/ml	2Standard product number	150Mlof3Number of standard products added150MlStandard diluent
3.75pg/ml	1Standard product number	150Mlof2Number of standard products added150MlStandard diluent

Sample addition: Set up blank wells (blank control wells without sample or enzyme-linked immunosorbent assay, all other steps are the same), standard wellsSample hole to be testedAccurately add standard samples on the enzyme-linked immunosorbent assay (ELISA) coated plate50MlAdd sample diluent to the test sample well first40MlThen add the sample to be tested10MlThe final dilution of the sample is5Double). Add the sample to the bottom of the enzyme-linked immunosorbent assay plate well, avoiding contact with the well wall as much as possible,gentlyshakemix well.

Incubation: Seal the plate with a sealing film and place it on the back37Incubate at 3 ℃0minute.

Liquid preparation: will30Concentrated washing solution with distilled water30Dilute twice before use

Washing: Carefully remove the sealing film and discard the liquid,spin-dryFill each hole with detergent and let it stand30Abandon in seconds, repeat like this5Next time, shoot dry.

Enzyme addition: Add enzyme labeled reagent to each well50MlExcept for blank holes.

Incubation: same operation3.

Washing: same operation5.

Color development: Add color developer to each hole firstA50MlAdd color developer againB50MlGently shake and mix well,37Color development in the dark for 10 minutes at ℃

Termination: Add termination for each holeliquid50μlTerminate the reaction(At this moment, blue turns to yellow).

Measurement: Zero with blank hole,450nmMeasure the wavelengths of each hole in sequenceabsorb lightDegree（ODValue)The measurement should be conducted after adding the termination solution15Conducted within minutes.

Summary of operating procedures:

calculate

Using the concentration of the standard substance as the horizontal axis,ODThe value is on the vertical axis, and a standard curve is drawn on the coordinate paper based on the sampleODFind the corresponding concentration from the standard curve; Multiply againdilution factor; or using the concentration of standard substances andODCalculate the linear regression equation of the standard curve based on the value, and calculate the sample'sODSubstitute the value into the equation, calculate the sample concentration, and then multiply it bydilution factor, which is the actual concentration of the sample.

Precautions

1The reagent kit should be taken out of the refrigerated environment and equilibrated at room temperature15-30Minutes later, the enzyme label coated plate can be used. If it is not used up after opening, the Flat noodles should be stored in a sealed bag.

2．Concentrated detergent solutionpossiblethere will becrystallizationPrecipitation can be dissolved by heating in a water bath during dilutionWashing does not affect the results.

3Each step of sample addition should use a sampler and its accuracy should be regularly checked to avoid experimental errors. It is best to control the sampling time at once5Within minutes, if there are a large number of specimens, it is recommended to use a sampling gun to add samples.

4.Please make a standard curve at the same time as each measurement, preferably with a double hole. If the content of the substance to be tested in the specimen is too high (sample)ODValue greater than the first hole of the standard holeODPlease dilute the sample diluent to a certain multiple first（nMeasure again after doubling, please calculate lastmultiplied byTotal dilution factor（×n×5）。

5.The sealing film is only for one-time use to avoid cross contamination.

6Please store the substrate away from light.

7Strictly 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.

8All samples, detergents, and various waste materials should be treated as infectious agents.

9Components from different batch numbers of this reagent must not be mixed.

Storage conditions and expiration date

1．Storage of reagent kit:；2-8℃.

2Validity period:6month