I would be grateful for some advice on how to detect % of antigen-specific B-cells after immunisation. Background: I have mice which express truncated IgM. After immunisation these mice produce no specific IgM antibodies but a normal amount of specific Ig. The question is whether specific IgM is not detectable in serum because it is monomeric and if IgM positive spleen cells (or other cells?) can be visualised by staining with antigen and FACS analysis. Approach: Ideally I would like to immunise these mice with a fluorescinated antigen (which one is best to elicit an extensive response?) and then gate the IgM positive cells and assess what percentage expresses the antigen. Thanks for help. marianne.bruggemann@bbsrc.ac.uk
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