Hi all,
I am planning on NP-imunisation of mice to study B cells. However, when looking at suppliers, there are various ratio's available. Which one is recommended, or what is the rationale to order one or the other? E.g. NP:CGG 7, or 26, or even >40?
Best,
Chris
Hi Christoph Matti
Unfortunately, there is no straightforward answer to your question, as it largely depends on the mouse strains used for your experiments and your final readout for the characterization of the B cell response.
If you fancy memory B cells a NP:CGG ratio > 26 is a good starting point as per Mark Shlomchik's work below. Please see the attached paper for more details and future reference.
Article CD80 and PD-L2 define functionally distinct memory B cell su...
For a quick characterization of the B cell response/ B cell compartment check out:
Article MYSM1-dependent checkpoints in B cell lineage differentiatio...
I hope that helps!
All the best & good luck with your experiments,
Michael
It is recommended to use CGG or other protein with a high number of NP for the induction of broad and robust B cells response (Low and high affinity B cell clones). For the anaysis of the B cell response you need to order a conjugate with a different protein and different loads of NP. For example, if you immunized your mice with NP-CGG (40 NP molecules), you need to use NP-BSA conjugate, in order to detect only the antibodies specific for NP. If you use a BSA conjugate with high load of NP, you will detect high and low affinity B cells clones specific for NP. But if you use a BSA conjugate with low NP (7), you will detect the high affinity clone, which show indirect evidence of selection and affinity maturation in the germinal centers.
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