Hi, I am looking for a specific anti-mouse IgG antibody for flow cytometry, but they all seem to cross-react (either with IgM or light chains of other immunoglobulins). I would like to look at IgG switched cells, but all the antibodies and protocols that I've found so far are for different isotypes (IgG1, IgG2a/b, IgG3). Shall I just combine all the antibodies? Anyone has done such experiment and can suggest an antibody (antibodies)?. Thanks in advance!
Hi Teja, what about an antibody against constant region of the light chain? I think its conserved in all subclasses.
Good luck!
Two comments/ideas
1. I have used Rat anti-mouse IgG1 (clone RMG1-1) with really good success in flow when looking at the % of GC B cells that expressed this class-switched heavy chain. This mAb does not cross-react with IgM, IgG3, IgG2a/c or IgG2b
2. How about taking a negative gating approach and using a mAb against Mouse Igµ (mAb b7-6 for example) to identify B cells that express BCR other than IgM. Using this approach, I would identify B cells as B220+ CD19+ CD79a (or b)+ and Igµ NEG. I am aware of the caveat that B cells expressing surface IgE or IgA BCR will be included in the Igµ NEG population. If you were really only concerned with IgG BCR+ B cells, you could include anti-IgA and anti-IgE mAb (heavy chain specific) to "exclude" these rare B cells as well.
Hi Greg, thank you for the suggestions. I already have IgA and IgM in my panel, I might include also IgE as you suggested.
Cheers, Teja
I have used the following IgG1 and IgG2a,b antibodies from BD. Works well in flow cytometry on B cells (no binding to IgM-positive cells). I usually use them in combination but they also work separately
IgG1-FITC (clone A85-1) BD #553443
IgG2a,b-FITC (clone R2-40) BD #553399
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