Biolegend
APC-Cy7 anti-mouse IgM
406516
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Antibodies are produced by plasma cells (PC) that are terminally differentiated B cells and play crucial roles in host protection against infections but are considered to mediate autoimmunity in patients with rheumatic diseases, such as systemic lupus (SLE) or rheumatoid arthritis (RA). Our research aims at understanding the biology of antibody secreting cells (ASC) in health and disease. This experiment aimed at measuring the amount of intracellular IgM expressed by CD138hi cells in naive mice.
Flow Cytometry
Mouse spleen
Surface staining with CD138 BV421, 20 min at 4C
Fc block (2.4G2 clone)
After permeabilization with cytofix/cytoperm anti-IgM APC-Cy7 1/5000 in Perm wash buffer, 20 min 4 C
N/A
Flow cytometer A5 Symphony BD
Around 40% of the CD138hi cells expressed intracellular IgM.
This antibody also stains surface IgM (in that case use at 1/150)
Cheap
Spread
It is a valid option to stain intracellular IgM