Problems that have already occurred in use:
1. Some customers did not read the instructions carefully. When using QuickAntibody adjuvant, they were emulsified with antigen according to the method of Freund's adjuvant. They were placed in a refrigerator at 4 degrees overnight, and the direct immunization was taken on the second day. The result was that the titer was not satisfactory.
Answer: QuickAntibody adjuvant does not require emulsification, and whether the adjuvant itself is mixed with the antigen, it will easily precipitate after standing for a long time. Therefore, it needs to be shaken before it can be formulated and injected.
2. Some customers did not read the instructions carefully. Before using QuickAntibody adjuvant, it was found that there was a precipitate in the adjuvant. It was thought to be a contaminant and was removed by centrifugation, resulting in an unsatisfactory titer of the experimental results.
Answer: The precipitation of QuickAntibody adjuvant is normal. Just shake it before use. If it is removed by centrifugation, it will affect the immune effect.
3. When a customer uses QuickAntibody adjuvant to immunize the hind leg calf muscle of the mouse, the ankle joint is mistaken as a knee joint, so that the immune site is too low, and a large amount of liquid flows out when 100 ul of adjuvant and antigen mixture is immunized .
Answer: The thighs of the hind legs of the mice are generally wrapped in the skin of the abdomen. The muscles of the hind legs that we normally see are the calf muscles. In addition, if the immune joint is immunized, because the immune site is far away from the effective immune response site, the antigen is easily degraded before it spreads to the lymph node, resulting in a decrease in the efficiency of the immune response.
4. Some customers use QuickAntibody adjuvant to immunize mice. Because there are more animals, the immunization time is longer. Finally, the adjuvant precipitates in the tube wall in the needle tube, resulting in higher antibody titer of the previously immunized mice. Antibody titers are low.
Answer: QuickAntibody adjuvant is prone to precipitation. It is recommended to use a small amount of adjuvant and antigen mixture to immunize each time before using it (two or three mice are better). After the operation is proficient, you can increase the immunity. The number of animals (eg, five or more mice per immunization).
5 , a customer using QuickAntibody adjuvant to prepare monoclonal antibody, the antigen is a polypeptide-coupled carrier protein, the ELISA titer against the polypeptide after immunizing two needles is only 1:200 , but successfully merged four hybridoma cell lines, two of which The produced monoclonal antibodies can be immunohistochemically and fully achieve their expected results!
Answer: The titer in our specification is the titer after immunization with protein antigen. For peptides and small molecules, it is impossible to achieve such a high titer after immunization! The reason is that peptides and small molecules are haptens, which have only one epitope or less than one epitope, and protein antigens have hundreds or thousands of epitopes. Customers are advised to test the ELISA titer against the carrier protein . If the titer range of the specification is reached and a certain level of peptide or small molecule titer is detected, it indicates that the immunization is successful and the subsequent fusion work can be continued. At this time, the titer of the polypeptide or small molecule depends on the immunogenicity and coupling efficiency of the epitope, and is basically independent of the adjuvant! However, the positive cloning rate of a polypeptide or small molecule monoclonal antibody is much lower than that of a protein antigen, so customers need to screen for more clones.
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