The results of the ELISA kit will be analyzed like this - Database & Sql Blog Articles

A few days ago, our salesperson received an online conversation from Mr. Qian, a teacher at Shanghai Jiao Tong University. Teacher Qian said that the test results of the elisa kit were not satisfactory and I didn't know what went wrong. Teacher Qian said: I synthesized an 8 amino acid antigen short peptide, and immunized the animal after coupling with BSA. After 1 month, the antibody titer against the antigen short peptide was determined by ELISA. The specific steps were: dissolving the antigen with carbonate buffer. After the short peptide, the plate was wrapped in a 4 degree refrigerator for 18 hours, and then blocked with 1% OVA at 37 degrees for 2 hours, and the plate was washed 3 times. The serum to be tested was diluted with 10% calf serum, incubated at 37 degrees for 1 hour, washed 3 times, added secondary antibody, incubated at 37 degrees for 40 minutes, washed plate 3 times, added TMB color, the results were quite different, The negative control also developed color, and the OD value and the positive difference were not significant. What is the reason? Is it a problem with washing?
In response to this, Shanghai Jinma technicians responded that the ELISA kit results are not ideal for analysis based on these aspects:
1. Whether the washing plate is a string of holes, such as string holes, it may cause pollution; you should preferably wash the plate with more than 5 times for each primary and secondary antibody.
2. The problem of blocking solution, you said that the negative control is also colored, the most important problem should be closed, should be reconsidered whether it is suitable to use OVA closure, if the OVA closure effect is not good, you can choose other blocking solution. There are a lot of related posts in the selection of the blocking solution. You can check it out.
3. It is recommended that the ELISA kit be best served at 37 degrees for two hours, which may be better.
Do you understand all of the above? Shanghai Jinma provides customers with a full range of services. If you have any questions, please feel free to contact us.