E. coli cell lysis for Protein Isolation URGENT HELP PLEASE
Dr Mick Jones
m.d.jones at ic.ac.uk
Fri Jan 14 11:23:08 EST 2000
I am running a student practical next week, and I no longer have access
to a sonicator for making cell lysates from E. coli.
The experiment simply involves growing E. coli containing a pGEX fusion
construct, lysing the cells, and analysing the cleared lysate on an
SDS/PAGE gel and staining with Coomassie blue. We also use the lysate
to run on another SDS/PAGE and western blot and probe with a monoclonal
antibody against the GST protein.
The experiments work very well.
This time round I do not have access toa sonicator to lyse the E. coli
I usually work with a 2 ml culture of E. coli.
CAN ANYBODY HELP?
What I need is a simple method for lysing the cells that doesn't use a
It would be nice if it is simply a chemical disruption with relatively
simple lab raegents, as opposed to a commercial reagent.
The proteins we are looking at are soluble.
Any help and advice, protocols greatfully accepted
Please reply directly to my email address: m.d.jones at ic.ac.uk
Lecturer in Molecular Virology
Department of Infectious Diseases and Microbiology
Imperial College School of Medicine
Hammersmith Hospital, Du Cane Road, London, W12 0NN
Tel: 020-8383-3328; Fax: 020-8383-3394
(Note new London code)
email: m.d.jones at ic.ac.uk
"To Infinity and Beyond" Buzz Lightyear (Toy Story)
"Crackin' Toast, Gromit" Wallace (The Wrong Trousers)
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