Brain-infiltrating Cells Isolation and Analysis

Brain-infiltrating cells were prepared following our previously published protocol73 (link). Cells were blocked with unconjugated anti-CD16/32 (clone 93; BioLegend), then surface labeled with PerCP-conjugated anti-CD45 (BD Pharmingen, clone 30-F11), FITC-conjugated anti-CD11b (BD Pharmingen, clone M1/70), and PE-conjugated anti-Ly6C/G (Gr1; BD Pharmingen, clone RB6-8C5) or anti-Ly6G (BD Pharmingen, clone 1A8) at 1:200 dilution for 30 min at 4 °C. Flow cytometric analysis was performed on an Accuri C6 and gates were applied as previously described6 (link). Data were subsequently analyzed in FlowJo and SigmaPlot (Systat Software).

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Howe C.L., LaFrance-Corey R.G., Mirchia K., Sauer B.M., McGovern R.M., Reid J.M, & Buenz E.J. (2016). Neuroprotection mediated by inhibition of calpain during acute viral encephalitis. Scientific Reports, 6, 28699.

Publication 2016

PerCP-conjugated anti-CD45 FITC-conjugated anti-CD11b anti-Ly6G SigmaPlot

Brain Cd11b Cells Clone Dilution Fitc Flow cytometric

Corresponding Organization :

Other organizations : Mayo Clinic, WinnMed

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Variable analysis

independent variables

Unconjugated anti-CD16/32 (clone 93; BioLegend)
PerCP-conjugated anti-CD45 (BD Pharmingen, clone 30-F11)
FITC-conjugated anti-CD11b (BD Pharmingen, clone M1/70)
PE-conjugated anti-Ly6C/G (Gr1; BD Pharmingen, clone RB6-8C5) or anti-Ly6G (BD Pharmingen, clone 1A8)

dependent variables

Flow cytometric analysis

control variables

Cells were blocked with unconjugated anti-CD16/32 (clone 93; BioLegend)
Cells were surface labeled at 1:200 dilution for 30 min at 4 °C

controls

No positive or negative controls were explicitly mentioned.

Annotations

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