Shanghai Hengyuan and your dialysis ELISA assay for the determination of antibodies - Database & Sql Blog Articles

Recently, Zhou students from Nanjing Normal University called to inquire about the Elisa assay for antibody determination. Many customers may have some understanding of this problem. So let us the technical experts of Shanghai Hengyuan Biotechnology help us today. Do some simple dialysis:
The basic principle of the principle can be summarized as: combining the specific antigen-antibody immunological reaction with the enzymatic catalytic reaction to show the primary immunological reaction by the amplification of the enzymatic reaction. Therefore, enzyme immunoassay generally consists of two parts, one or several immunological reactions and one enzymatic reaction. During the specific binding of the antigen and the antibody, an enzyme conjugate, such as an enzyme-labeled anti-antibody, is added, and then the substrate of the enzyme is added to generate a colored product under the enzyme catalysis, reflecting the amount of the antigen or antibody to be assayed. Operating procedures, types and characteristics of the operational procedures: detection of antibodies using indirect elisa. The antigen was coated with the purified antigen, and the antibody was blocked with an unrelated protein for at least 2 hours at 4 ° C overnight. The sample to be tested was added, and the sample was incubated for 1 hour at room temperature. After washing, the enzyme-labeled antibody was added, and then incubated at room temperature and washed to obtain a substrate. Color development, color development to a certain extent to terminate the reaction, read the results on the microplate reader or judge with the naked eye. Type: There are many types of elisa specific methods, and the standardization of reagents has not been finally solved. The incubation time, temperature, type of washing liquid and number of washings are all factors influencing the determination. However, with reference to the above basic process, no matter how it changes, it is inseparable from the antigen-antibody reaction, the immunological reaction of the enzyme-labeled antibody, and the enzyme catalysis. Features: 1. High sensitivity, detectable concentration at ug~mg/ml level; 2. Strong specificity; 3. Good repeatability; 4. Fast, simple and inexpensive. Experimental conditions Solid phase carrier: There are many substances that can be used as a solid phase carrier, such as cellulose, polyacrylamide, polystyrene, polypropylene, cross-linked dextran, glass, silicone rubber, agarose gel, and the like. Can appear in various forms; test tubes, micro-reaction plate recesses, beads ..... The most commonly used is polystyrene products, which have the best adsorption properties. The mechanism of solid phase coating is generally nothing more than physical adsorption and covalent cross-linking. Blocking: occupying uncoated voids on the carrier with unrelated proteins. The commonly used blocking solution is 1% bovine serum albumin or 10% calf serum. . Samples to be tested: The dilution should be appropriate. If the concentration is too large, non-specific adsorption will cause false positives. Excessive dilution will affect the immunological reaction between antigen-antibody. The optimum dilution is selected, and the dilution of the optical density in the experimental system is preferably about 1.0. Enzyme conjugates: The use of high quality enzyme conjugates is key to the success of elisa. Precautions The standardization of reagents for enzyme-linked immunoassay has not been fully resolved. The time, temperature, type of washing solution and number of washings in the incubation process are all factors that influence the final determination. In order to obtain satisfactory results in the experiment, it should be done during the operation. 1. Positive and negative controls must be set for each test. 2, the closure should be complete. 3. Washing should be determined.

Shanghai Hengyuan Biotechnology has been operating elisa kit for more than ten years and has been providing customers with a full range of services. After purchasing the kit, if there are any problems during the experiment, our technicians will provide technical guidance and collect the customer. Some of the problems encountered in the sample, that is, pre-sale, sale, and after-sales can provide technical services. Therefore, choosing Hengyuan products will definitely make you feel safe and use it more smoothly!