Bovine IgG, ≤1 mg/mL
Hemoglobin, ≤20 mg/dL
cell culture | hybridoma: suitable
cell culture | insect: suitable
cell culture | mammalian: suitable
≤10 EU/mL endotoxin
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At this time, Sigma-Aldrich does not offer delipidated bovine serum (either FBS or calf serum).A method for lipid depletion, though not necessarily that of Sigma:1. While mixing, gradually add 10 g fumed silica (Product S5130 or Aerosil) per liter of serum2. Mix at room temperature and then transfer to 2-8 °C for overnight mixing.3. After overnight mixing, remove flumed silica (Aerosil) by centrifugation at 3000-5000 rpm for 5 minutes.4. Retain the supernatant. The lipids should stick to the silica. There may be some protein loss
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.
Once Fetal Bovine Serum has been thawed, the bottle can be used in cell culture for up to 7 days for most cell types when stored refrigerated at 2-8°C. If the cells are not fastidous (e.g. HeLa or fibroblasts), the thawed and refrigerated serum can be used for up to 30 days. For some extremely fastidious cell lines, it may be necessary to screen individual lots for acceptable performance. For uses such as blocking for various immuno procedures, usage will generally be for 30 days and perhaps somewhat longer when stored at 2-8°C. For other procedures, usage beyond 30 days storage at 2-8°C will need to be determined.
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