Author: Brabb, Thea; von Dassow, Peter; Ordonez, Nadia; Schnabel, Bryan; Duke, Blythe; Goverman, Joan
Title: In Situ Tolerance within the Central Nervous System as a Mechanism for Preventing Autoimmunity Document date: 2000_9_18
ID: kcygxo7h_7
Snippet: Flow Cytometry. Single-cell suspensions (10 6 ) of splenocytes, LN cells, or CNS cells (10 5 ) were resuspended in 10 l of whole mouse serum and anti-CD16/CD32 (0.05 g/ml) for 15 min at room temperature (RT). Samples were then incubated for 30 min at RT in the dark with primary antibodies diluted in 50 l of FACS ® staining buffer (FSB [0.05% sodium azide and 5% FCS; Atlanta Biologicals] in PBS). Cells were washed, resuspended in 50 l of secondar.....
Document: Flow Cytometry. Single-cell suspensions (10 6 ) of splenocytes, LN cells, or CNS cells (10 5 ) were resuspended in 10 l of whole mouse serum and anti-CD16/CD32 (0.05 g/ml) for 15 min at room temperature (RT). Samples were then incubated for 30 min at RT in the dark with primary antibodies diluted in 50 l of FACS ® staining buffer (FSB [0.05% sodium azide and 5% FCS; Atlanta Biologicals] in PBS). Cells were washed, resuspended in 50 l of secondary antibody diluted in FSB, and incubated for 20 min at RT. Cells were washed and resuspended in either 300 l of FSB or fixed in FSB with 0.37% formaldehyde. Cells (at least 3 ϫ 10 5 per sample) were analyzed with a FACScan™ flow cytometer (Becton Dickinson). Fluorescence is plotted on a log scale.
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