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#heat inactivation

2 public questions tagged with this topic.

Animal Tissue Culture 1

Excessive heat inactivation may damage:

Thermal denaturation affects serum constituents differentially according to thermodynamic stability determined by disulfide content, hydrophobic core packing, and glycosylation. Growth factors platelet-derived growth factor BB homodimer stabilized by two interchain disulfides plus intrachain cysteines, fibroblast growth factor basic bFGF all beta-sheet trefoil without disulfides relatively labile, transforming growth factor beta dimer with nine disulfides but still aggregation prone above fifty degrees, and epidermal growth factor small 6 kDa with three disulfides but beta sheet conformation s

Ref: Brunner 2010 FBS heat damage growth factors oxidation PDGF; Freshney excessive heat albumin denaturation reduces cloning efficiency.

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Animal Tissue Culture 1

Heat inactivation of serum is performed at:

Freshly collected fetal bovine serum contains intact complement system cascade components of innate immunity capable of opsonization, chemotaxis, and direct lysis. Components include classical pathway C1 complex C1q six headed collagen-like recognizing immune aggregates, C1r C1s serine proteases cleaving C4 C2 forming C3 convertase C4b2b, alternative pathway factor B D, lectin pathway mannose binding lectin MBL associated serine proteases MASP, central protein C3 three chain alpha beta gamma 185 kDa convertible to active C3b depositing via thioester covalent attachment onto cell surface hydrox

Ref: Gibco FBS heat inactivation 56°C 30 min complement C2 denaturation; Abbas Immunology complement C1q classical pathway removal.

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