FBS Heat Inactivated — 56°C 30 min Complement-Depleted | Cell Culture | SeamlessBio
FBS Heat Inactivated — 56°C / 30 min | Complement-Depleted
Heat inactivation at 56°C for 30 minutes destroys the complement system in FBS — a cascade of ~30 serum proteins that can lyse cells, activate immune responses, and interfere with certain cell culture and immunological applications. While heat inactivation is not required for most standard cell lines, it is essential for complement-sensitive primary cells, immune cell assays, and specific virological applications.
Important note: Heat inactivation is not recommended as a default for all applications. The process reduces some growth factor levels (PDGF, IGF-1) and denatures certain proteins. Only use heat inactivated FBS when complement activity is specifically problematic for your application. For standard immortalised cell lines, regular FBS is preferred.
When Heat Inactivation is Required
| Application | Why Complement Must be Removed |
|---|
| Primary haematopoietic cell culture | Complement directly lyses red blood cells and certain white blood cells — heat inactivation prevents non-specific cell death in PBMC and bone marrow cultures |
| ELISpot assays (T cell, B cell) | Active complement in FBS causes background cell lysis that elevates spot counts and reduces assay specificity |
| Mixed lymphocyte reaction (MLR) | Complement can activate lymphocytes non-specifically — must be removed for clean alloresponse measurements |
| Viral infection assays | Complement can neutralise enveloped viruses via the classical pathway — heat inactivation prevents viral neutralisation in infection assays |
| Cell lines with Fc receptors | Complement activation via FcR on macrophages and NK cells creates background cytotoxicity |
| Hybridoma culture (some protocols) | Complement can lyse hybridoma cells expressing surface immunoglobulin — heat inactivation sometimes specified in hybridoma protocols |
What Heat Inactivation Does and Does Not Do
| Parameter | Effect of Heat Inactivation (56°C/30 min) |
|---|
| Complement proteins (C1q–C9) | ✅ Destroyed — all complement cascade components inactivated |
| IgG, IgM antibodies | Partially reduced — some denaturation occurs |
| Albumin, transferrin | ✅ Largely preserved |
| EGF, FGF | ✅ Largely preserved |
| PDGF, IGF-1 | ⚠️ Some reduction — heat labile growth factors affected |
| Mycoplasma, viruses | ⚠️ NOT fully inactivated — heat inactivation is not a sterilisation step |
| Endotoxin | ❌ NOT reduced — endotoxin is heat-stable |
Specifications
| Parameter | Specification |
|---|
| Heat inactivation | 56°C for 30 minutes — validated complement destruction |
| Complement activity | Not detectable post-inactivation (haemolytic assay) |
| Sterile filtration | 0.1 µm post-inactivation |
| Endotoxin | Tested per lot (LAL) |
| Mycoplasma | Tested — negative |
| Origin | EU and/or US |
| Volumes | 100 mL, 500 mL | Bulk on request |
| Documentation | CoA (incl. complement inactivation validation), CoO, MSDS |
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