Travel Perfume Spray Bottle,Plastic Perfume Spray Bottles,Empty Perfume Spray Bottles,Refillable Perfume Spray Bottle GuangZhou ShenRuo Plastic Manfacturing CO.,Ltd , https://www.cnbottle.com
Instruction Manual of Human Interleukin 1β (IL-1β) Kit
Human interleukin 1β (IL-1β) enzyme-linked immunoassay Purpose: This kit is used to determine the content of interleukin 1β (IL-1β) in human serum, saliva and related liquid samples. Experimental principle This kit uses the double antibody sandwich method to determine the level of human interleukin 1β (IL-1β) in the specimen. The microplate was coated with purified human interleukin 1β (IL-1β) antibody to prepare a solid-phase antibody. Interleukin 1β (IL-1β) was added to the microwells coated with mAb in turn, and then labeled with HRP The interleukin 1β (IL-1β) antibody binds 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 interleukin 1β (IL-1β) in the sample. The absorbance (OD value) was measured with a microplate reader at a wavelength of 450 nm, and the concentration of human interleukin 1β (IL-1β) in the sample was calculated by a standard curve. Operation steps Dilution of standard products: This kit provides one original standard product. The user can perform dilution in a small test tube according to the following chart. 40pg / ml No. 5 standard 150μl original standard added 150μl standard dilution 20pg / ml No. 4 standard 150μl standard 5 added 150μl standard dilution 10pg / ml No. 3 standard 150μl No. 4 Add 150μl standard dilution 5pg / ml No. 2 standard 150μl No. 3 standard add 150μl standard dilution 2.5pg / ml No. 1 standard 150μl No. 2 standard add 150μl standard dilution Add sample: respectively Set blank wells (the blank control wells do not add samples and enzyme-labeled reagents, the rest of the steps are the same), standard wells, and sample wells to be tested. Accurately add 50μl of the standard on the enzyme-coated plate, add 40μl of sample diluent to the sample well, and then add 10μl of the sample to be tested (the final dilution of the sample is 5 times) Add the sample and add the sample to the bottom of the well of the microplate, try not to touch the wall of the well, shake gently to mix. Incubation: Seal the plate with a sealing film and incubate at 37 ° C for 30 minutes. Solution: Dilute the 30-fold concentrated washing solution with distilled water 30-fold and wash it later: Carefully remove the sealing film, discard the liquid, spin dry, fill each well with the washing solution, let it sit for 30 seconds, then discard, repeat 5 Time, pat dry. Enzyme addition: Add 50μl of enzyme label reagent to each well, except for blank wells. Incubation: The operation is the same as 3. Washing: The operation is the same as 5. Color development: add 50μl of developer A to each well, then add 50μl of developer B, mix gently, and develop at 37 ° C in the dark for 15 minutes. Termination: add 50μl of stopper solution to each well to stop the reaction (blue at this time) Li to yellow). Measurement: The absorbance (OD value) of each well was measured in sequence with the blank air conditioner at zero and 450 nm wavelength. The measurement should be carried out within 15 minutes after adding the stop solution. Calculate the standard concentration as the abscissa and the OD value as the ordinate. Draw a standard curve on the coordinate paper, and find the corresponding concentration from the standard curve according to the OD value of the sample; multiply by the dilution factor; The linear regression equation of the standard curve is calculated from the concentration and the OD value, the OD value of the sample is substituted into the equation, the sample concentration is calculated, and then multiplied by the dilution factor, which is the actual concentration of the sample. Matters needing attention 1. The kit should be equilibrated at room temperature for 15-30 minutes before being taken out of the refrigerated environment. If the enzyme label coated plate is unopened, the strip should be stored in a sealed bag. 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. 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 (the OD value of the sample is greater than the OD value of the first well of the standard well), please first dilute it with a certain multiple (n times) of the sample diluent and then determine it. When calculating, please multiply the total dilution Multiple (× n × 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 based on the reading of 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 it is different from the English manual, the English manual shall prevail.