What is heat inactivated serum?
What is Heat Inactivation? The practice of heat inactivating serum was originally developed when only serum from adult animals was available for cell culture. Adult serum contains various immune factors, particularly serum complement, which may inhibit or destroy cells under certain conditions.
What is serum complement inactivation?
Human complement is inactivated by plasmin, the proteolytic enzyme of plasma or serum active at or near neutrality. The addition of streptokinase to human serum, which converts plasminogen to plasmin, also causes the inactivation of complement components C’2 and C’4 and varying amounts of C’1.
What does heat inactivated mean?
Definition. Heat Inactivation (HI) – a process by which serum is maintained at a temperature of 56± 2° for 30± 2 minutes. Equipment. Water bath capable of maintaining temperatures of 56± 2°C. Preferably a shaking water bath.
Does heat inactivation destroy antibodies?
(2) Heating sera may destroy the advanced structure of the antibody, which may interfere with the binding to the assay; however, heating treatment could also inactivate the complement system in sera, which could avoid non-specific reaction and decrease the background in immunological assays [26], [27].
Why is serum inactivation important?
Heating serum at 56 degrees is used to inactivate complement in several immunological assays. During heating, both heat-labile and heat-stable anticomplementary activity (ACA) develop. While heat-labile ACA can be completely inactivated, heat-stable ACA increases progressively with continued heating.
Is it necessary to heat inactivate serum?
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 heat inactivate serum?
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. Swirl the serum every 5 to 10 minutes to ensure uniform heating and to prevent protein coagulation at the bottom of the bottle.
Why is heat inactivation important?
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.
What does heat inactivation do to viruses?
Heat treatment is a widely-used decontamination method, notably used for viruses [10]. It is thought to inactivate viruses principally by denaturing the secondary structures of proteins and other molecules, resulting in impaired molecular function [11].
Should I heat inactivate FBS?
Since we have changed to fetal bovine serum, we find that heat inactivation is not necessary for most cell lines.
Why do we need to heat inactivate FBS?
Heat inactivation of FBS degrade active phase protein in particular Degrading complement proteins which can effect the outcome of the experiment and immune response in general.
What is the purpose of heat inactivated FBS?
Heat Inactivation – Serum is inactivated by raising the temperature to 56 °C for 30 minutes under controlled conditions. The objective of heat inactivation is to destroy complement activity in the serum without affecting the growth-promoting characteristics of the product.
Why is serum heated?
Are all FBS heat inactivated?
All lots of FBS – Premium are manufactured in our ISO 9001:2015 certified facility. Heat Inactivation – Serum is inactivated by raising the temperature to 56 °C for 30 minutes under controlled conditions.
What happens if you heat inactivate FBS for too long?
Notes: Heating the serum for longer than 30 minutes or higher than 56°C will have an adverse effect on the efficacy of the serum and will most likely cause an increase in the amount of cryo-precipitate that might form.