chloroform in colony blots
Jeff E. Janes
jejanes at silibone.cchem.berkeley.edu
Wed Nov 25 14:21:25 EST 1998
Hi all,
I'm doing some experiments in which I lift colonies onto
nitrocellulose, then visual for a specific protein using
antibody-AP conjugate. Some of the protocols I have seen
(none of which seem quite satisfactory for me) call for treating
the nitrocellulose-bound cells (E. coli) to chloroform vapors
for 15 minutes prior to lysozyme treatment. Does anyone know
the purpose of this step?
Also, I would prefer to blot for periplamsic proteins only,
but all protocols indicate complete cell lysis. Does anyone
know of a protocol which will work for this?
Thanks,
Jeff
More information about the Methods
mailing list