Why heat inactivated serum? Is it necessary to do heat inactivation?
1.Why heat inactivated serum? Is it necessary to do heat inactivation?
56 ℃ heating can inactivate the complement system. Activated complement is involved in lysing cellular events, stimulating smooth muscle contraction, releasing histamine from cells and platelets, and activating lymphocytes and macrophages. In immunological studies, ES cell culture, insect cells and smooth muscle cells, heat-inactivated serum is recommended.
Experiments have shown that heat-inactivated serum, correctly processed, is not required for most cells. The treated sera had little or no effect on the growth of the cells and often resulted in a decrease in the rate of cell growth even though the quality of the sera was usually affected by the high-temperature treatment. The heat-treated serum, the formation of sediment will be significantly increased, the deposit observed under an inverted microscope like “black spots”, often make researchers mistaken for serum contamination, and the serum on the 37 ℃ In the environment, this sediment will be more and more, so that the researchers mistakenly believe that the division of microbial expansion.
2.How to thaw serum will not make the product quality damage?
When thawing serum, follow the recommended gradual thawing method (-20 ° C to 4 ° C to room temperature). If the temperature of thawing is too high (e.g., -20 ° C to 37 ° C) when thawing, experiments show that sedimentation is very likely to occur. Thaw serum, please shake it at any time, so that the temperature and composition uniformity, reduce the occurrence of precipitation. Do not put the plasma at 37 ℃ for too long, if left at 37 ℃ for too long, the serum will become cloudy, while many of the more unstable components of serum will, therefore, be compromised, and affect the quality of the serum. Serum heat inactivation is very likely to cause increased sediment, if not necessary; you can do this step without having to do. If you must do the heat inactivation of serum, please follow the principle of 56 ℃, 30 minutes, and shake evenly at any time. Temperature is too high, too long or uneven shaking, will cause increased sediment.
3.serum thawed flocculent precipitate appeared, how to deal with?
Precipitation in serum occurs for a number of reasons, but the most common cause is due to the denaturation of lipoproteins in the serum, and fibrin, one of the proteins that form the blood clot, is also present in the serum after thawing of the serum, Is also one of the main causes of sediment. However, these flocculent sediments do not affect the quality of the plasma itself. If you want to remove this flocculent sediment, the serum can be aliquoted into sterile centrifuge tubes and centrifuged at 400g. The supernatant can then be filtered into the medium. We do not recommend that you filter out these flocculent sediments because it may clog your filter membrane.
4 What is the best way to save your serum?
The recommended serum should be stored at -5 ° C to -20 ° C. If stored at four ℃, please do not exceed one month. If you can not run out of a bottle, it is advisable to sterilize the aliquots to sterile centrifuge tubes and return to freezing.
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