monoclonal antibody immunization schedule - (Mar/27/2012 )

hi im having a problem in determining the concentration and volume for the soluble antigen that im going to raise in mice using an ip technique. The concentration that im planning to use is 250microgram/ml. Can anyone guide me on this?my immunization schedule is every two weeks interval. What would be the appropriate volume for antigen for each immunization?

Your immunizations can be anywhere from 10ug/mouse to 200ug/mouse. I usually use anywhere from 25 to 100ug/mouse. It may depend on how much antigen you have available. If you are lucky someone has already done this immunization and wrote a paper about it. If this is the case study that paper thouroughly and decide your injection volume. One last note, a mouse Can take alot by IP injection so 200uL of mixed antigen and ajuvant can be inject fairly easily. Hope this helps.

The previous answer is good, to had a bit of complement most of the available adjuvant (Sigma or MPL TDM or even Immun Easy) are at a maximum of 1 to 1 mix with your Antigen so if you inject 200microliters of your mixture, you can go up to 100microliters of your protein. We routinely use with success 40 micrograms of protein/immunisation .

You can eventually use IP and subcutaneous to be complementary !