human male plasma (Type AB)
hemoglobin, ≤20 mg/dL
cell culture | mammalian: suitable
≤10 EU/mL endotoxin
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10 - Combustible liquids
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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.
Product H6914 is prepared from whole blood that is allowed to clot, centrifuged and the serum removed. There is no additional processing required.Product H4522 is prepared from anticoagulated (sodium citrate) whole blood. This is centrifuged to pellet the cells (RBC, WBC, some platelets) and the plasma is removed. This plasma is then treated with calcium to activate the clotting cascade. The preparation is centrifuged again and the supernatant is removed. This product can also be called plasma derived serum.Product H6914 will contain more growth factors since it is allowed to spontaneously clot and there will be the release of growth factors from many of the cells (WBC, platelets) present. The cells are removed before the plasma is clotted for Product H4522, so this plasma will have fewer growth factors present (more similar to what is circulating). Unless testing for growth factors, most researchers are looking for Product H6914.We offer both products heat inactivated.Product H5667 Human Serum Heat Inactivated, from male AB clotted whole bloodProduct H3667 Human Serum Heat Inactivated, from human male AB plasma
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