Rows of square glass bottles with white caps are placed on a metallic surface. The bottles contain a reddish-brown liquid and are labeled with text. The setting resembles a laboratory or scientific environment.

Heat Inactivation of Serum Products

The objective of Fetal Bovine Serum heat inactivation, or inactivation of any other serum, is to destroy complement activity in the serum without affecting the growth-promoting characteristics of the product. Removal of complement activity from serum, such as fetal bovine serum, is not required for most cell cultures but may be necessary for cultures that are sensitive to the complement activity. Since heat inactivation of the serum may, to some extent, decrease the growth performance properties of the serum, this procedure should only be performed if required for optimal cell growth. Researchers should evaluate the applicability of heat inactivation regarding their own application.

Preparation

  1. Thaw the desired Serum until at room temperature, if thawed in refrigerator allow serum to set at room temperature.

  2. Heat agitating water bath (set at 56°C) and fill with water to ensure bottles will be immersed to cover the level of the serum.

  3. Gently mix the contents of the bottle/serum thoroughly before placing in the water bath.

  4. It is recommended to have a weight to secure each bottle and prevent tipping while water bath is agitating.
Bottles of fetal bovine serum in a laboratory setting are arranged in rows. The bottles have white caps and labels. Some bottles have visible frothy bubbles inside.
Bottles of fetal bovine serum are arranged in rows. Each bottle has a label with a lot number and a "heat inactivated" sticker. The bottles are capped with white lids and contain a brown liquid.

Heat Inactivation

  1. Once water bath temperature reaches 56± 2°C, place the serum bottles in the water bath (include control bottle with thermometer to monitor internal temperature of serum).

  2. Set the water bath to agitate or swirl the bottles every 10 minutes to ensure serum remains uniform. NOTE: Agitation prevents gelling and will promote a uniform heating.

  3. When the control bottle reaches an internal temperature of 56± 2°C, begin timer for 30 minutes.

  4. After 30 minutes, remove the bottles and transfer into cooling rack/space. Cool the bottles for at least 1 hour before aliquoting or return to freezer.

Frequently Asked Questions

 

What is FBS heat inactivation?
Heat inactivation is a process that involves heating FBS to deactivate complement proteins without compromising the serum's growth-promoting properties.

When Is Heat Inactivation of FBS Necessary?
Heat inactivation aims to destroy complement activity in the serum without impacting its growth-promoting properties. While most cell cultures don't require the removal of complement activity, it is essential for cultures sensitive to it.

How is FBS heat inactivation performed?
The standard method involves heating the FBS at 56°C for 30 minutes while regularly mixing to ensure uniform heating.

Does heat inactivation affect the quality of FBS?
Properly performed heat inactivation should not affect the growth-promoting properties of FBS. However, improper technique can degrade some of the serum's beneficial components.

What are the alternatives to heat inactivation?
Alternatives include using commercially available pre-inactivated FBS or other treatment methods like filtration to remove complement activity.

For more commonly asked questions, go to our FAQ page.

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