2 week induced CK-p25 mice and age-matched controls were perfused with 50 mls PBS to wash away blood and minimize macrophage contamination in the brains. Hippocampal tissue was harvested immediately after perfusion and a single-cell suspension was prepared as described previously56 (link). Fluorescence-activated cell sorting (FACS) was then used to purify CD11b+ CD45low microglia cells using APC conjugated CD11b mouse clone M1/70.15.11.5 (Miltenyi Biotec, #130-098-088) and PE conjugated CD45 antibody (BD Pharmingen, #553081). Cells were collected directly into RNA lysis buffer (Qiagen, #74104).
Protocol detail
Purification of Microglia from CK-p25 Mice
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Corresponding Organization :
Other organizations : Massachusetts Institute of Technology, Broad Institute, Harvard University
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Variable analysis
independent variables
- Induction of CK-p25 in mice
- Microglia cells purified using FACS
- Age-matched control mice
- Positive control: Age-matched control mice
- Negative control: Not explicitly mentioned
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