How do you heat inactivate fetal bovine serum?
Heat Inactivation – Serum is inactivated by raising the temperature to 56 °C for 30 minutes under controlled conditions.
Why do we heat inactivated fetal bovine serum?
A common treatment of FBS is heat-inactivation, where FBS is heated at 56°C for 30 minutes in a water bath with occasional shaking. The purpose is to inactivate whatever components of the complement system are present in the FBS , and other potential unknown inhibitors of cell growth.
Is heat inactivation of FBS necessary?
In past years, Coriell used all heat-inactivated serum for its cell cultures to inactivate the complement protein found in newborn calf serum. Since we have changed to fetal bovine serum, we find that heat inactivation is not necessary for most cell lines.
How do you activate fetal bovine serum?
Technical Tips 1 – Heat Inactivation of Fetal Bovine Serum
- Thaw the serum slowly to 37°C and mix the contents of the bottle thoroughly.
- Place the thawed bottle of serum into a 56°C water bath containing enough water to immerse the bottle to just above the level of the serum.
- Begin timing for 30 minutes.
How is complement inactivated?
Human complement is inactivated by plasmin, the proteolytic enzyme of plasma or serum active at or near neutrality. The inactivation of complement by the addition of streptokinase to human serum is inhibited by high hydrogen ion concentrations, low temperature, and elevated ionic strength.
What is the difference between fetal bovine serum and fetal calf serum?
There is no difference between FBS and FCS, it is the same serum with name preference. However, the ‘F’ for fetal is very important, and catalog numbers from different companies are critical for cell culture.
What is heat inactivation?
Heat inactivation is a convenient method for stopping a restriction endonuclease reaction. Incubation at 65°C for 20 minutes inactivates the majority of restriction endonucleases that have an optimal incubation temperature of 37°C.
How do you heat inactivate human serum?
Swirl the bottles every 3-5 minutes to ensure uniform heating of the serum. Monitor the temperature of the control bottle closely and begin timing when the temperature reaches 56±2°C. After 30 minutes (25 minutes for equine/human serum), remove the serum bottles and cool slowly to room temperature.
How do you heat inactivate?
How to heat inactivate serum:
- Thaw the serum slowly (see above).
- Prepare a water bath large enough to accommodate the number of bottles being treated.
- Place bottles of thawed serum in the water bath.
- Bottles should be agitated gently during the heat inactivation process to ensure uniform heating throughout the serum.
Why do we inactivate complement?
The objective of heat inactivation is to destroy complement activity in the serum without affecting the growth-promoting characteristics of the product. Removal of complement activity from the serum is not required for most cell cultures, but may be necessary for cultures that are sensitive to the complement activity.
Where does fetal bovine serum come from?
Fetal bovine serum (FBS) is a common component of animal cell culture media. It is harvested from bovine fetuses taken from pregnant cows during slaughter.
What is the purpose of fetal bovine serum?
Fetal bovine serum (FBS) is a byproduct of harvesting cattle for the meatpacking industry—it’s used extensively by both academic and industrial researchers as a supplement to basal growth medium in cell culture applications. FBS is the liquid portion that remains after blood is drawn from bovine fetus coagulates.