Nov 09, 2024

Public workspaceHeat Inactivate FBS

 Forked from Heat Inactivate FBS
DOI
dx.doi.org/10.17504/protocols.io.j8nlk9eewv5r/v1
  • 1Microbiology and Parasitology Department, University of Navarra
  • Paolo Edoardo Ginatta Arosemena: Edited and simplified protocol for our laboratory needs
  • Parasitolab
Paolo Edoardo Ginatta Arosemena
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DOI: dx.doi.org/10.17504/protocols.io.j8nlk9eewv5r/v1
External link: http://www.serumsourceintl.com/pdf/heat_inactivation_instructions.pdf
Protocol Citation: Paolo Edoardo Ginatta Arosemena 2024. Heat Inactivate FBS. protocols.io https://dx.doi.org/10.17504/protocols.io.j8nlk9eewv5r/v1
Manuscript citation:
Based largely from http://www.serumsourceintl.com/pdf/heat_inactivation_instructions.pdf
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: November 08, 2024
Last Modified: November 09, 2024
Protocol Integer ID: 111797
Keywords: FBS Heat Inactivate, Inactivation,
Abstract
Heat inactivate FBS to denature compliment. Critical for trypanosomes but not for EA.hy 926 cells.
Before start
Thaw FBS in the fridge.
Thaw
Thaw
Allow serum to acclimate at room temperature for a minimum of 10 minutes or refrigerate overnight at 2° C to 8° C.
The serum may then be completely thawed at Temperature37 °C in a water bath. Swirl the bottles throughout the thawing process every 10-15 minutes as this will disperse the released salts and proteins uniformly.

Prepare a water bath, if not already done so during the thawing process, that can be temperature controlled to Temperature56 °C in for the heat inactivation process. There must be enough water to submerge the entire contents of all bottles.
Heat-inactivate
Heat-inactivate
30m
30m
Place the serum into the Temperature56 °C in water bath for Duration00:30:00

30m
Gently swirl the bottles every 3-5 minutes to make certain the serum remains uniform throughout heating.
Remove the heat inactivated serum after the 30 minutes and gently swirl once again.
In cell culture hood, aliquot 50 mls into 10 50 ml containers.
Label as Heat Inactivated FBS
Causes of Failure
Causes of Failure
Improper Thawing
Inactivating serum that has not been thoroughly mixed homogeneously
Inactivating serum above the recommended temperature
Inactivating serum for a longer period of time than recommended 30 minutes