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发表于 2011-10-31 13:08:49
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找到一篇早期文章了：http://onlinelibrary.wiley.com/doi/10.1002/eji.200526189/abstract
全文我看不到，但在一篇引用它的文章中看到了简要步骤，似乎利用0.5％吐温：
In brief, fresh PBMCs were washed in PBS, fixed in 1 ml of PBS with 1% paraformaldehyde and 0.05% Tween, and kept overnight at 4°C. Cells were treated twice with 0.5 ml of DNase at 100 Kunitz/ml according to manufacturer’s instructions (Sigma-Aldrich). Cells were incubated with mouse anti-human Foxp3 (clone150D/E4, a kind gift from Alison Banham, University of Oxford, Oxford, United Kingdom) for 1 hour at room temperature and washed with FACS buffer (PBS, 3% FCS, 0.5% Tween-20, and 0.05% azide). Foxp3 staining was detected by staining with Alexa Fluor 488 goat anti-mouse IgG (Molecular Probes; Invitrogen). Cell-surface staining was performed using anti-CD4–PerCP (BD) and anti-CD25–PE (Dako) for 20 minutes at room temperature. |
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发表于 2011-10-27 16:38:52
