Interleukin 10 (IL-10) ELISA kit Calculate the concentration of human interleukin 10 (IL-10) in the sample by the standard curve. Introduction: Both human and mouse IL-10 genes are located on chromosome 1, whose genome includes 5 exons and 4 introns, and It can have the binding position of NF-κB and AP-1. Human and mouse IL-10 have 81% and DNA and amino acid levels, respectively. 73% homology. DNA sequence analysis showed that IL-10 and EB virus genome open reading frame region â… (BCRF- â… ) have 70% homology. Some people call the gene product of BCRF-I as virus IL-10 (vIL-10), suggesting that EBV may be And IL-10 may become an anti-inflammatory treatment by promoting the expression of IL-1ra by monocytes. Tests show that IL-10 can effectively avoid the death caused by LPS-induced mouse shock. Kit composition: The supernatant should be centrifuged again if precipitation occurs during storage. Afterwards, 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 sterile tubes and centrifuged for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully and save the process If a precipitate forms, it should be centrifuged again. Pleural and ascites, cerebrospinal fluid reference implementation. 4. Supernatant of cell culture: When detecting secreted components, collect with a sterile tube. Centrifuge for about 20 minutes (2000-3000 rpm /Minute). Collect the supernatant carefully. When detecting the intracellular components, dilute the cell suspension with PBS (PH7.2-7.4), the cells The concentration reaches about 1 million / ml. Through repeated freezing and thawing, the cells are destroyed and the intracellular components are released. Centrifuge for 20 minutes Around clock (2000-3000 rpm). Collect the supernatant carefully. If a precipitate forms during storage, it should be centrifuged again. 5. Organize the specimen: weigh the weight after cutting the specimen. Add a certain amount of PBS, PH7.4. Quick frozen storage with liquid nitrogen spare. After the specimen melts, it still maintains a temperature of 2-8 ° C. Add a certain amount of PBS (PH7.4), use hand or homogenate The device homogenizes the specimen sufficiently. Centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. After serving To be tested, the rest are frozen for use. Steps Standard 100μl, then add standard dilution 50μl in the first and second wells, mix well; then from the first well, the first Take 100μl from each of the two wells and add them to the third and fourth wells respectively, then add standard dilutions to the third and fourth wells respectively 50μl, mix well; then take 50μl each in the third and fourth wells and discard, then take 50μl each and add to the fifth and In the sixth well, add 50ul of standard dilution solution to the fifth and sixth wells respectively, mix well; Take 50μl from each well and add it to the seventh and eighth wells respectively, then add 50μl of standard dilution solution to the seventh and eighth wells, After mixing, take 50μl from the seventh and eighth wells and add them to the ninth and tenth wells. 50μl of quasi-product dilution, mix well and take 50μl from the ninth and tenth wells and discard. (After dilution, the volume of each well is 50μl, The concentrations are 240μg / L, 160μg / L, 80μg / L, 40μg / L, 20μg / L). 2. Add sample: set up blank wells separately (the blank control wells do not add samples and enzyme reagents, the rest of the steps are the same), to be tested Sample hole. Add 40μl of sample diluent to the test sample well of the enzyme-coated plate, and then add 10μl of the test sample (The final dilution of the sample is 5 times). Add the sample to the bottom of the well of the microplate, try not to touch the wall of the well, shake gently Mix thoroughly. 3. Incubation: seal the plate with the sealing film and incubate at 37 ° C for 30 minutes. 4. Mixing solution: Dilute 20 times concentrated washing solution with distilled water 20 times and reserve. 5. Washing: Carefully peel off the sealing film, discard the liquid, spin dry, fill each well with washing liquid, let stand for 30 seconds and then discard, so Repeat 5 times and pat dry. 6. Add enzyme: add 50μl of enzyme label reagent to each well, except blank well. 7. Incubation: The operation is the same as 3. 8. Washing: The operation is the same as 5. 9. Color development: add 50μl of developer A to each well, and then add 50μl of developer B, mix gently, and develop color at 37 ℃ in the dark 15 minutes. 10. Termination: Add 50μl of stop solution to each well to stop the reaction (at this time the blue will turn to yellow). 11. Determination: Measure the absorbance (OD value) of each well in sequence with the blank air conditioner at zero and 450nm wavelength. Determination should be terminated Within 15 minutes after the solution. Precautions: After use, the slats should be stored in sealed bags. 2. Crystals may be precipitated in the concentrated washing liquid, which can be heated and dissolved in a water bath during dilution, and the results will not be affected during washing. 3. The sampler should be used at each step of sample addition, and the accuracy should be regularly checked to avoid test errors. It is best to control the sampling time within 5 minutes. If there are many specimens, it is recommended to use a volley gun to add samples. 4. Please make a standard curve at the same time of each measurement, it is best to make a double hole. If the content of the test substance in the specimen is too high (sample OD value Is greater than the OD value of the first well of the standard well), please dilute it with a certain multiple (n times) of the sample diluent before measuring. When calculating, please multiply the total dilution factor (× n × 5). 5. The sealing film is limited to one-time use to avoid cross-contamination. 6. Please keep the substrate away from light. 7. Strictly follow the instructions, and the test results must be determined by the microplate reader. 8. All samples, washing liquids and various wastes should be treated as infectious agents. 9. The components of different batches of this reagent shall not be mixed. 10. If there is any difference with the English manual, the English manual shall prevail. Calculation: Taking the concentration of the standard as the abscissa, the OD value is the ordinate, Draw a standard curve on coordinate paper, according to the OD of the sample The value is determined by the standard curve; then multiplied by the dilution Multiple; or calculate the standard using the concentration and OD value of the standard The linear regression equation of the quasi-curve, the OD value of the sample Substitute into the equation, calculate the sample concentration, and multiply by the dilution The multiple is the actual concentration of the sample. Kit performance: 2. The batch and approval shall be less than 9% and 11% respectively Forehead Pdo Thread,Pdo Thread Lift Neck, Pdo Threads Nasolabial Folds,Pdo Smooth Threads Shijiazhuang Asa Technology Co., Ltd. , https://www.hskinlift.com
user's Guide
This reagent is for research use only
Purpose: This kit is used to determine human serum, plasma and related
The content of interleukin 10 (IL-10) in the liquid sample.
Experimental principle:
This kit uses the double antibody sandwich method to determine the level of human interleukin 10 (IL-10) in the specimen. Coat the microplate with purified human interleukin 10 antibody to make a solid-phase antibody, add interleukin 10 to the monoclonal antibody-coated microwells in turn, and then combine with HRP-labeled goat anti-porcine antibody to form the antibody- Antigen-enzyme-labeled antibody complex, after thorough washing, added substrate TMB 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 IL-10 in the sample. Measure the absorbance (OD value) at 450nm with a microplate reader,
Ingested the mammalian IL-10 gene for their own survival. IL-10 antagonists may have anti-EB virus
Kit composition 48-well configuration 96-well configuration Description
Enzyme label coated plate 1 × 48 1 × 96 2-8 ℃
Sealing film 2 pieces (48) 2 pieces (96)
Standard product: 360μg / L 1 bottle × 0.5ml 1 bottle × 0.5ml 2-8 ℃
Standard dilution 1 bottle × 1.5ml 1 bottle × 1.5ml Store at 2-8 ℃
Enzyme label reagent 1 bottle × 3 ml 1 bottle × 6 ml Store at 2-8 ℃
Sample diluent 1 bottle × 3 ml 1 bottle × 6 ml Store at 2-8 ℃
Developer A solution 1 bottle × 3 ml 1 bottle × 6 ml 2-8 ℃
Developer B Solution 1 bottle × 3 ml 1 bottle × 6 ml 2-8 ℃
20 times concentrated washing solution 1 bottle × 30ml 1 bottle × 50ml Store at 2-8 ℃
Stop solution 1 bottle × 3ml 1 bottle × 6 ml Store at 2-8 ℃
Instructions 1 copy 1 copy
Sample processing and requirements:
1. Serum: room temperature blood coagulates naturally for 10-20 minutes, centrifuged for about 20 minutes (2000-3000 rpm). Collect carefully
2. Plasma: EDTA, sodium citrate or heparin should be selected as anticoagulant according to the requirements of the specimen, mixed for 10-20 minutes
1. 1. Dilution and sample addition of standard products: 10 standard wells are set on the enzyme-coated plate, and the first and second wells are added respectively
1. The kit should be equilibrated at room temperature for 15-30 minutes before being taken out of the refrigerated environment.
1. The correlation coefficient R between the linear regression of the sample and the expected concentration is more than 0.990.
Detection range: storage conditions and validity period
10μg / L -360μg / L 1． Kit storage :; 2-8 ℃.
2. Validity: 6 months
Interleukin 10 (IL-10) ELISA kit instructions