What are the benefits and disadvantages of using heat-inactivated FBS in cell culture?
- Heating inactivates complement. Active complement can participate in cytolytic events, contract smooth muscle, release histamine from mast cells and platelets, and activate lymphocytic and macrophage cells. Applications where heat-inactivated serum is recommended include immunological studies and culturing of embryonic stem cells (ESCs), insect cells, and smooth muscle cells.
- Heat inactivation helps to achieve bottle-to-bottle and lot-to-lot stability by neutralizing many factors that can vary largely from lot to lot.
- There aren't necessarily disadvantages to heat inactivation of FBS, but there is some evidence that suggests there may be no added benefit to it unless you are carrying out immune studies.
Note: Heat inactivation is performed in a 56 degrees C water bath for 30 min with swirling every 10 min or so for heat distribution and to lower the degree of protein aggregation/flocculant precipitation. Note: If the time or temperature is exceeded, the serum may thicken to a gel. If this occurs, the serum is no longer usable. Unnecessary heat inactivation can take up time and potentially lead to wasted reagents if a mistake is made during the protocol1.
1. Pellerin, et al., Bioengineering, published in 2021.
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How much of the total protein measured in Fetal Bovine Serum (heat inactivated or otherwise) is attributed to albumin?
Our CoAs only capture the total protein content not specifically the amount attributed to albumin.
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How much Fetal Bovine Serum (FBS) do I need to add to my medium?
FBS is added to culture medium at a concentration of 2-10% to provide attachment factors, nutrients, and hormones for mammalian cells, as well as to be a buffer against disruptions like pH changes and endotoxins. FBS has significant amounts of embryonic growth promoting factors like hormones, carrier proteins, and macromolecular proteins. It also has low levels of antibodies and other growth-inhibiting components. For most basal medium, 10% FBS is used. Some applications require up to 20% supplementation, so researchers need to determine what is optimal for their specific application and cell line.
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How long can I store Gibco Fetal Bovine Serum at 4 degrees C?
This product can be stored at 4 degrees C for up to 4 weeks. Once the medium has been supplemented with serum, we recommend using it within 2-4 weeks.
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How should I thaw Gibco Fetal Bovine Serum?
We recommend thawing the serum overnight at 4 degrees C or in a 37 degrees C water bath, removing as soon as it is thawed. Once thawed, aliquot into single-use sizes and freeze the aliquots. Each aliquot should ideally be thawed only one additional time as repeated freeze-thaw cycles are not recommended.
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