Paula Lavery wrote:
>> I would like to immunize mice with plasmid DNA in order to get monoclonal
> antibody. In order to get the best response of I have to injure the muscle
> with cardiotoxine 5 days prior to DNA injection. I do not have the great
> success with cardiotoxine injection. Death rate of mice is more than 50%.
> The concentration of the cariotoxine is given to mice 100 micromolar in
> PBS, 100 microliter bilateral injection. If you can suggest some advice, it
> would be helpful.
I tried 50ul 0.25% bupivicaine bilaterally, followed by 50ug
plasmid/flank the following day. Two rounds of this and I got a decent
serum response. Had to follow this up with a dose of a soluble form of
the protein (it was a cell surface protein and I suspect I needed to get
the antigen to the spleen rather than draining lymph nodes but that's
only a guess). This worked very well but of course it was just one
attempt with one antigen and in one strain of mouse (balb/c's).
Hope this helps
Clive Tregaskes
Institute for Animal Health, Compton, UK
