BD Biosciences
Alexa Fluor 647 Rat Anti-Mouse CD162
562806
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The goal of this research study was to determine if B cell depletion altered effector T cell formation during LCMV infection.
Flow Cytometry
Mouse Lymph Node Leukocytes
Wash cells once in FACS buffer (PBS plus 1% BSA or serum). Block Fc receptors and stain for viability in PBS for 20 minutes on ice.
Fc block (TruStain FcX)
Wash cells once in FACS buffer. Stain with isotype control or PSGL1 antibody (1:500 dilution, 100 uL per million cells) along with other cell surface-directed antibodies for 20 minutes on ice. Wash cells once in FACS buffer. Resuspend cells in 100-250 uL PBS containing 1.5% paraformaldehyde.
N/A
Flow Cytometer
PSGL-1 clearly segregated pre-memory (PSGL-1+Ly6C-) from effector (PSGL-1+Ly6C+) and follicular helper (PSGL-1loLy6Clo) T cells. With B cell depletion by CD20 mAb, there is a substantial decrease in the proportion of pre-memory T cells.
10.4049/jimmunol.1302848
Clearly distinguishes PSGL-1 expression.
Could be a brighter antibody.
A robust method to subset effector T cell populations.