Heat inactivating a 50ml serum aliquot - 56degC for how long? (Mar/03/2006 )

Hihi,

How long should the duration be for inactivating serum if it is in a 50ml falcon tube aliquot?

Anyone have experience doing this?

My lab does not heat inactivate their sera and i only need to make 1/2 a litre of growth medium. The serum are already stored in 50ml falcon tube aliquots. I dont think 30mins is recommended for such a small amount of serum ... anyone know?

30' too

1. Thaw the serum slowly to 37°C and mix the contents of the bottle thoroughly.

2. 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.

3. 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.

4. After 30 minutes at 56°C cool the serum immediately, preferrably in an ice bath.

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. For volumes less than 500 ml of serum to be heat inactivated, reduce the time of incubation in the 56°C water bath.

I thaw my serum for roughly 24 hrs on 4°C. Then heat inactivate for 30' on 56°C with swirling every 10 minutes and put back on 4°C.

Works very fine.

erm .. maybe i did not make myself clear. I know that the standard heat inactivation process for 500ml stock of serum is 56degC for 30minutes.

However I am only intending to heat inactivate a 50ml aliquot of serum (currently stored in a falcon tube). For volumes less than 500 ml of serum to be heat inactivated, the time of incubation has to be reduced to avoid deactivating the serum components which will result in poor cell cultures.

I dont know what is the optimum incubation time for 50ml serum stored in plastic.

Anyone successfully done this? thanks in advance

hi,
fred33's answer was sweet and simple. if you recollect little bit of physics knowledge.
in heat inactivation procedure we should give 56C temperature for 30 minutes to the serum, it does not matter whether serume is in 50ml falcon tube or it is in 1.5ml tube.
just matter of incubation time rather volumes.

gud luck