In this video, we describe a method to study the organelle dynamics in B lymphocytes following their activation with antigen-coated microbeads.
Protocol
NOTE: The following steps were performed using IIA1.6 B cells. 1. B cell activation with antigen-coated beads Preparation of antigen-coated beads To activate B cells, use NH2-beads covalently coated with antigen (Ag-coated beads), which are prepared using 50 µL (~20 x 106 beads) of 3 µm NH2-beads with activating (BCR-ligand+) or non-activating (BCR-ligand-) antigens. <li…
Divulgations
The authors have nothing to disclose.
Materials
IIA1.6 (A20 variant) mouse B-lymphoma cells
ATCC
TIB-208
Murine B-cell lymphoma of Balb/c origin that expresses an IgG-containing BCR on its surface without FcγIIR