Sigma EGF was tested in Craig Coopersmith’s lab to verify its basic safety in the mouse. This was performed given that the older EGF product was discontinued and new EGF (E4127; 203EUR/157USD per 0.1mg) was purchased; we will all use it for pRCT.
6 CD-1 mice (6 weeks old) per group used (3F & 3M). EGF diluted in saline given twice/day for three days at the concentration of 75ug/kg.
We dilute EGF (E4127, 0.1mg) in 0.5ml saline to make a stock solution, with a concentration of 200ug/ml. For long-term storage, aliquot it into pre-chilled sterilized microcentrifuge tubes and immediately snap-freeze the vials in liquid nitrogen prior to re-storage at -70°C. Repeat freeze-thaw cycles should be avoided. As such, the aliquotes are expected to be stable for 6 months to a year.
Each day, a new frozen stock EGF tube is thawed up and tubes for injections are prepared for that day.
The EGF stock of 200ug/ml is very concentrated, so when applying EGF to mice, it needs to be diluted about 100 times, of course, depending on the weight of mice.
The above and additional details are contained in the SOP.