This reagent is for research purposes only: this kit is used to determine the content of interleukin 2 (IL-2) in human serum, plasma and related liquid samples.

Experimental principle:

This kit uses the double antibody sandwich method to determine the specimen

(IL-2)) level. Microporous plates were coated with purified chicken interleukin-2 (IL-2) antibody to prepare solid-phase antibodies, and chicken interleukin-2 (IL-2) was added to the monoclonal antibody-coated microwells in turn, and then It is combined with HRP-labeled goat anti-chicken antibody to form an antibody-antigen-enzyme-labeled antibody complex. After thorough washing, the substrate TMB is added for color development. TMB is converted into blue under the catalysis of HRP enzyme, and into the final yellow under the action of acid. The color depth is positively correlated with chicken interleukin-2 (IL-2) in the sample. The absorbance (OD value) was measured with a microplate reader at a wavelength of 450 nm, and the content of chicken interleukin-2 (IL-2) in the sample was calculated by a standard curve.

Kit composition:

The kit consists of 48 well configurations and 96 well configurations.

Instructions 1 copy 1 copy

2 pieces of sealing film (48) 2 pieces (96)

One sealed bag

Enzyme label coated plate 1 & time48 1 & time96 2-8 ℃

Standard product: 360ng / L 0.5ml & time1 bottle 0.5ml & time1 bottle Store at 2-8 ℃

Standard diluent 1.5ml & time1 bottle 1.5ml & time1 bottle Store at 2-8 ℃

Enzyme label reagent 3 ml & time1 bottle 6 ml & time1 bottle Store at 2-8 ℃

Sample diluent 3 ml & time1 bottle 6 ml & time1 bottle Store at 2-8 ℃

Developer A solution 3 ml & time1 bottle 6 ml & time1 bottle Store at 2-8 ℃

Developer B Solution 3 ml & time1 bottle 6 ml & time1 bottle Store at 2-8 ℃

Stop solution 3ml & time1 bottle 6ml & time1 bottle Store at 2-8 ℃

20 times concentrated washing solution 20ml & time1 bottle 23ml & time1 bottle 2-8 ℃

Sample processing and requirements:

1. Serum: room temperature blood coagulates naturally for 10-20 minutes, centrifuged for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully and centrifuge again if a precipitate appears during storage.

2. Plasma: EDTA or sodium citrate should be selected as anticoagulant according to the requirements of the specimen. After mixing for 10-20 minutes, centrifuge for about 20 minutes (2000-3000 rpm) Collect the supernatant carefully. If a precipitate forms during storage, it should be centrifuged again.

3. Urine: collected in a sterile tube and centrifuged for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. If a precipitate forms during storage, centrifuge again. Pleural and ascites, cerebrospinal fluid reference implementation.

4. Cell culture supernatant: When detecting secreted components, collect with a sterile tube. Centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. When detecting the components in the cells, dilute the cell suspension with (PH7.2-7.4), and the cell concentration will reach about 1 million / ml. Through repeated freezing and thawing, the cells are destroyed and the intracellular components are released. Centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. If a precipitate forms during storage, it should be centrifuged again.

5. Organize the specimen: after cutting the specimen, weigh it. Add a certain amount, PH7.4. Quickly freeze and save with liquid nitrogen for later use. After the specimen melts, it still maintains a temperature of 2-8 ° C. Add a certain amount (PH7.4) and homogenize the specimen by hand or homogenizer. Centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. After aliquoting, a portion is to be tested, and the rest is frozen for future use.

6. The specimen should be extracted as soon as possible after collection. The extraction should be carried out according to relevant literature. The experiment should be carried out as soon as possible after extraction. If the test cannot be performed immediately, the specimen can be stored at -20 ℃, but repeated freezing and thawing should be avoided.

7. The sample containing NaN3 cannot be detected because NaN3 inhibits the activity of horseradish peroxidase (HRP).

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