Heat inactivation of serum is a topic of interest to many trainers. The expensive serum contains precious substances such as growth factors, vitamins, etc., and it is not necessary to put them at a temperature above 50 ℃ for 30 minutes. of.
Nevertheless, the heat inactivation of serum in most laboratories is still performed as a routine. Most experimenters do not consider the negative effects of heat treatment on growth factors, amino acids and other components in serum, in our technical hotline The most frequently mentioned is whether the serum should be thermally inactivated. Below we will discuss and explain the thermal inactivation of fetal bovine serum.
The purpose of heat inactivation is to remove heat-sensitive substances such as complement in serum, but inactivation of complement in fetal bovine serum is obviously unnecessary. Triglia and Linscott once measured the complement components in commercial fetal bovine serum. They found that C1 and C6 contained in fetal bovine serum only reached 1-3% of the adult animal serum, while other complement components were only 5-50 of the adult animal. %, As the main component of complement C3 in fetal bovine serum is almost impossible to detect. Through the complement fixation experiment, we have also obtained similar results in multiple different batches of fetal bovine serum. Even in undiluted serum, no obvious hemolysis was found. In addition, the process of preheating the culture medium before cultivation in most laboratories also has an inactivating effect on the heat-sensitive complement.
According to the survey, at least 70% of the researchers inactivated the serum only because they followed the normal operation, or take it for granted. The recommended temperature for routine inactivation is between 45 ° C and 62 ° C, and the time varies from 15 minutes to 60 minutes. The most commonly used method is heat treatment at 56 ° C for 30 minutes. With the improvement of serum collection, processing, and processing technology, many of the reasons that were previously considered to be heat inactivation are no longer valid. Only a few researchers who performed heat inactivation on serum have confirmed the effectiveness and necessity of this step in experiments. Sex.
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