Hyclone A-2151-L bovine calf serum, defined, iron-supplemented, 500 ml bottles.
STERILE (do not filter)
WARNING: The serum is sensitive to light and the yellow plastic should be on the bottle or the room should be only dimly lit.
The serum bottles are stored in the -80 °C freezer. The night before aliquoting the serum into tubes, transfer the frozen bottle into the refrigerator, placing it on a plastic box top to keep it from cracking any surface in the refrigerator.
The next day, the serum may still be partially frozen. Heat the bottle until the serum melts in the 37 °C water bath, swirling every 2 minutes. When the serum has melted, add a second similarly sized bottle filled with ice cold water (with no ice) and holding a thermometer, and turn the water bath up to 56 °C.
When the thermometer reads over 54 °C, start a timer for 30 minutes. Swirl both bottles every 5 minutes. At the end of 30 minutes, put the serum bottle in ice water (with some ice) for 10 minutes, swirling every few minutes.
This is a good time to turn the water bath back down to 37 °C.
Aliquot the serum 9.1 ml into 13 ml tubes (an extra 0.1 ml is added since serum sticks to the walls and not all is recovered). For the last tube with a non-standard amount, just label it mentioning how many ml were added.
Put the tubes into the round paper boxes and put them into the -20 °C freezer to freeze overnight. (Putting them into racks or directly into the -80 °C freezer can cause the tubes to break). The next day, (or later) the tubes can be placed horizontally in the plastic boxes and stored in the -80 °C freezer (for long term storage) or the -20 °C freezer (for shorter term storage).
If this was the last bottle of serum, be sure to order more serum well in advance of the next time we will need more.
Copyright © 2002 Dr. Michael M. Segal, Department of Neurosurgery, Brigham & Women's Hospital, Boston MA 02115, USA. Many of the solutions are based on recipes from the Furshpan and Potter labs; the recipes given here are the versions we use now in the Segal lab. Individual copies may be made of this page for use in educational institutions.