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发表于 2012-3-3 10:38:48
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找到一篇做猪KC细胞的，给你做个参考，如果你做人的标本，需要注意一下，抗体不能订文章中的那些克隆，因为那些针对猪KC的。
下面是文章中的方法学部分摘节，我觉得很奇怪，他们用CD14、CD11b、CD18、CD45来标记KC细胞，但单核细胞也全部都表达这些表达，如何能够保证单核不被混进去？
附件中是全文PDF。
101313KsP.pdf
(342 KB, 下载次数: 43)
Flow Cytometry
KC were released from culture plates on day 3 using 12 mM lidocaine in PBS at 37°C (26). Cells were blocked with PBS containing 0.5% BSA, 0.02% sodium azide and stained with anti-CD11b (American Type Culture Collection, Manassas, VA, HB-249), anti-CD14 (R&D Systems, Minneapolis, MN), anti-CD18 (Developmental Studies Hybridoma Bank, Iowa City, IA, H52), and anti-CD45 (ABD Serotec, Raleigh, NC) for 1 hr followed by two washes with buffer. Mouse isotypes IgG1k and IgG2b (eBioscience, San Diego, CA)
were used as negative control. Propidium iodide staining was used to exclude dead cells. Goat anti-mouse IgG PE (Jackson ImmunoResearch Laboratories, West Grove, PA) was used as secondary. All flow cytometry data was collected and analyzed on the Accuri C6 flow cytometer and CFlow software (Accuri, Ann Arbor, MI).
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发表于 2012-3-2 22:32:11
我先去试一下
