Heat inactivation of serum is performed to inactivate complement (which can cause cell lysis or downstream problems in immunoassays). If there are immunoglobulins present in the serum that can bind to the cells in culture (either specifically or non-specifically), there is a chance for antibody-mediated complement cytotoxicity (ADCC, causing cell death). It may also reduce the titer of some mycoplasma and some viruses. Often, heat-inactivated serum is used because of the adoption of a previous protocol. It is mainly used when culturing hematopoetic cells or performing immunological type assays such as mixed-lymphocyte reactions or neutralization assays.FBS may not be heat inactivated as this is not required for all applications. It limits the loss of the other protein activity that may be minimally present in the serum.A protocol for heat inactivation:1. Allow serum containers to come to room temperature. Swirl bottles of serum immediately before adding to the water bath.2. A water-filled control bottles (T1) should be placed in a 56 °C water bath. Place the containers in the water bath up to the serum line. Do not completely submerge the containers. When the temperature of T1 reaches 56 °C, start the timer set for 30 minutes. Do not allow temperatures to exceed 56 °C. Higher temperatures will denature the proteins, gel the serum or alter the performance of the serum.3. Gently swirl the bottles every 5 to 10 minutes while in the water bath and check the temperature in the control bottle. It is not uncommon for plastic containers to bow slightly after heat treatment. This is not harmful to the serum.4. At the end of 30 minutes remove the serum bottles from the water bath. Aliquot the serum into smaller volumes and freeze. Sigma-Aldrich does not recommend repeated thaw-freeze cycles.
H1138
Horse Serum
Donor Herd, USA origin, Heat inactivated, sterile-filtered, suitable for cell culture
Synonym(s):
HS, equine sera, equine serum, horse sera, sera, serum
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About This Item
biological source
horse serum
Quality Segment
sterility
sterile-filtered
composition
Hemoglobin, ≤20 mg/dL
origin
USA origin
technique(s)
cell culture | mammalian: suitable
impurities
≤10 EU/mL Endotoxin
shipped in
dry ice
storage temp.
−20°C
Application
- as a media supplement in the differentiation medium to culture the pheochromocytoma (PC12) cells
- in immunocytochemistry and immunohistichemistry
- to study the therapeutic effect of serum treatment on cardiomyocyte viability
Preparation Note
Prepared by heating at 56 °C for 30 minutes.
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This Item | |||
|---|---|---|---|
| sterility sterile-filtered | sterility sterile-filtered | sterility sterile-filtered | sterility sterile-filtered |
| technique(s) cell culture | mammalian: suitable | technique(s) cell culture | hybridoma: suitable, cell culture | mammalian: suitable | technique(s) cell culture | hybridoma: suitable, cell culture | mammalian: suitable | technique(s) cell culture | mammalian: suitable |
| biological source horse serum | biological source horse serum | biological source horse serum | biological source sheep |
| origin USA origin | origin USA origin | origin USA origin | origin USA origin |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C | storage temp. −20°C |
| composition Hemoglobin, ≤20 mg/dL | composition hemoglobin, ≤20 mg/dL | composition Hemoglobin, ≤20 mg/dL | composition hemoglobin, ≤50 mg/dL |
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Storage Class
10 - Combustible liquids
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves
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Why is serum Heat-Inactivated (HI)?
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