My understanding is that the effect of cyclohexamide is reversible upon
removal of the agent and also that it could affect cellular events other
than the inhibition of protein synthesis. This information was passed down
to me and hence do not have a reference(s) for it. Whenever I have used
it, cyclohexamide was retained in all downstream sample processing steps in
at least one-half the concentration used to inhibit protein biosynthesis.
If reversibility is a problem, the use of 20--100 uM of emetine, whose
effect on protein synthesis is irreversible, should solve it. Emetine
sensitivity of the cell line under study should be determined empirically
because it varies widely. It is also toxic when used in higher
concentration than optimal or in optimal concentrations for prolonged
periods of time; this feature too is cell type dependent.
sebastian
Sebastian Joyce, Ph.D.
DEPARTMENT OF MICROBIOLOGY & IMMUNOLOGY
THE PENNSYLVANIA STATE UNIVERSITY COLLEGE OF MEDICINE
MILTON S.HERSHEY MEDICAL CENTER
500 UNIVERSITY DRIVE
HERSHEY, PA 17033-0850
Tel: 717-531-4163
Fax: 717-531-4600