We know that ELISA is a new technology in immunodiagnosis, and has been successfully applied to immunodiagnosis of infectious diseases, parasitic diseases and non-infectious diseases caused by various pathogenic microorganisms. It has also been applied to the quantitative determination of large-molecule antigens and small-molecule antigens. Based on the results used, the ELISA method is considered to be sensitive, specific, simple, fast, stable, and easy to operate automatically. Today, Shanghai Hengyuan Biological Technology Co., Ltd. came to talk about the operation steps and applications of the elisa kit. The steps of elisa kit: 1. Double antibody sandwich method 1. Coating: Dilute the antibody to a protein content of 1-10 μg / ml with 0.05MPH9. Carbonate coating buffer. Add 0.1ml to the reaction well of each polystyrene plate, overnight at 4 ℃. The next day, the solution in the well was discarded and washed 3 times with washing buffer for 3 minutes each time. (Referred to as washing, the same below). 2. Add sample: add 0.1ml of the diluted test sample to the above-mentioned coated wells and incubate at 37 ℃ for 1 hour. Then wash. (Make blank wells, negative control wells and positive control wells at the same time). 3. Add enzyme-labeled antibody: add 0.1ml of freshly diluted enzyme-labeled antibody (dilution after titration) to each reaction well. Incubate at 37 ° C for 0.5 to 1 hour and wash. 4. Add substrate to develop color: add 0.1ml of temporarily prepared TMB substrate solution to each reaction well at 37 ℃ for 10 to 30 minutes. 5. Terminate the reaction: Add 0.05ml of 2M sulfuric acid to each reaction well. 6. Judgment of results: The results can be directly observed with the naked eye on a white background: the darker the color of the reaction well, the stronger the positive degree, and the negative reaction is colorless or extremely light. According to the depth of the color, "+", "-" Indicates. OD value can also be measured: on ELISA detector, at 450nm (410nm if ABTS color is developed), the OD value of each well is measured after zero adjustment with a blank control well, if it is greater than 2.1 times the OD value of the specified negative control , That is positive. 2. Indirect method 1. Dilute the known antigen to 1 ～ 10μg / ml with coating buffer, add 0.1ml to each well, overnight at 4 ℃; 2. Wash 3 times the next day; 3. Add a certain dilution of the test sample (unknown antibody) 0.1ml to the above-mentioned coated wells, incubate at 37 ℃ for 1 hour, and wash; 4. (Do blank, negative and positive well control at the same time) Add 0.1ml fresh diluted enzyme-labeled secondary antibody (anti-antibody) to the reaction well; 5. Incubate for 35-60 minutes at 37 ° C, wash; 6. 'Last wash with DDW. The remaining steps are the same as 4, 5 and 6 of the "double antibody sandwich method". Application of elisa kit: 1. Immunoenzyme staining the location of various intracellular components. 2. Study the synthesis of anti-enzyme antibodies. 3. A trace of immunoprecipitation reaction appears. 4. Quantitative detection of antigen or antibody components in body fluids. The main agent products of Shanghai Hengyuan Biology: ELISA kit: ELISA kit, human ELISA kit, rat ELISA kit, pig ELISA kit, mouse ELISA kit, chicken, duck, monkey, rabbit ELISA kit. Imported medium: microbial medium, color medium, E. coli O157 medium, total bacteria medium, vibrio test medium, listeria test medium. Serum: fetal bovine serum, human serum, animal serum. Biological reagents: Amresco, Sigma, Spectrum imported reagents. Standard reference products: imported reference products, imported reference herbs, imported standard products, imported standard reagents. Our company adheres to the corporate culture of focusing on quality, keeping promises, active communication and innovative services, and actively participates in technological innovation and technical services in the biological field, and strives to contribute to China's scientific research. Shanghai Hengyuan Biological Technology Co., Ltd. launched a double 11 promotion campaign, loyally looking forward to your call to consult, I wish you good health and good luck in everything! Botulinum Toxin Type A,Botox Botulinum Toxin Type A,Botulinum Toxin Type A Injection,Botulax Botulinum Toxin Type A Yanbian Nabalu E-commerce Co., Ltd. , https://www.nabalubeauty.com