How Do You Inactivate Horse Serum?

Donor Horse Serum, Heat Inactivated is treated by heat inactivation where serum is exposed to a temperature of 56 °C for 30 minutes under controlled conditions. Serum products should be stored and handled correctly to assure long-term stability and to preserve growth performance consistency throughout its shelf-life.

Does horse serum need to be heat inactivated?

In general, it is not necessary to use heat-inactivated serum to culture most cell types. We recommend using heat-inactivated serum when working with immune type cells or for immunological applications. Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

How is serum inactivated?

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.

How do you heat inactivate calf 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.

How do you heat inactivate sera?

Perform heat inactivation at 56 °C for a period of 30 minutes by heating the laboratory-scale serum bottles in a water bath. During this process, stir the serum and shake it at least from time to time, while avoiding foaming.

Why do we need to heat inactivated serum?

Adult serum contains various immune factors, particularly serum complement, which may inhibit or destroy cells under certain conditions. Heating serum is intended to inactivate serum complement.

What is the difference between fetal bovine serum and horse serum?

Horse Serum contains more immunoglobulins than fetal bovine serum (FBS), has an increased protein content, and is a cost-effective alternative to fetal bovine serum. Horse Serum is used for diagnostic assays, supplementation during mycoplasma growth, or for cultivation of hematopoietic stem cells and neuronal cells.

Why do you heat-inactivate?

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 will you inactivate the complement components in a serum?

Abstract. 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 is inactive serum?

in·ac·ti·vat·ed se·rum. serum that has been heated to 56°C for 30 minutes to destroy the lytic activity of complement.

At what temperature do you heat inactivate FBS?

56± 2°C
The following protocol is suggested for the heat inactivation of fetal bovine serum ( FBS ). FBS is heated to 56± 2°C for 30± 2 minutes in agitating water-bath. Heat Inactivation (HI) – a process by which FBS is maintained at a temperature of 56± 2° for 30± 2 minutes.

What happens when serum is heated?

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.

Do I need to heat inactivate restriction enzyme?

Restriction enzymes are commonly inactivated by a heat treatment after digestion is complete. However, heat tolerance varies between enzymes, and in some cases is insufficient to completely inactivate particular restriction enzymes.

What are other ways to inactivate the complement?

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.

How do you heat inactivate FBS Gibco?

How do I heat-inactivate serum? Heat it at 56°C in a water bath for 30 minutes and swirl the bottle every 10 minutes or so. For accuracy, use a second bottle of similar size as a control and add an equivalent volume of water to the control bottle. Place a thermometer in the control bottle to see when 56°C is reached.

How do you inactivate an enzyme?

Heat treatment, or blanching, without doubt constitutes the simplest and most direct method of enzyme inactivation. It consists of brief immersion (from 1 to 6 min depending on size) of the product in water, boiling syrup, or steam close to 100 °C. Catechol oxidases are inhibited above approximately 70 °C.

Why must we heat inactivate fetal bovine serum FBS before adding it to our media preparation and using that preparation to culture mammalian cells?

Heat inactivation of serum (incubation at 56 °C for 30 minutes) can help to reduce the risk of contamination, since some viruses are inactivated by this process; however, this process also denatures some proteins and destroys nutrients in the serum.

Why are enzymes inactivated at high temperatures?

At high temperature, many enzymes are inactivated by aggregations at hydrophobic sites which are exposed on denaturation. Isolating denatured enzymes via hydrophobic interactions with other material is a significant method to prevent enzymes from aggregation.

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 [24], and other potential unknown inhibitors of cell growth.

Why is horse used in serum?

Horses have been deemed particularly useful for producing therapeutics for human use because of their relatively large blood (or urine) volume, which can be collected repeatedly and extracted for antibody, hormone, or other protein isolation, and because of their general ease in handling and maintenance [1].

What do you use horse serum for?

What Are the Uses of Horse Serum? A: Horse Serum, like other animal sera, can be used as a supplement to basal growth medium in cell culture. Horse Serum applications include diagnostic assay, animal and human vaccine productions and as a supplement in bacteriological media.