作者查詢 / tharaohfu
作者 tharaohfu 在 PTT [ Biotech ] 看板的留言(推文), 共33則
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5F推: 我們的做法是切片完後一樣冰-20,開始染色前先放到-2001/11 20:30
6F→: 度C的methanol,接著PBS wash再做block。01/11 20:30
4F推: 先確認挑出來的clone是否為你要的tumor cell06/28 23:41
5F→: 找個anti-human epithelial marker跑個flow馬上就知道06/28 23:42
6F→: 當然要先查一下paper H446會表現什麼marker06/28 23:45
7F→: 同步再確認mapping data有沒有什麼設定有錯誤06/28 23:47
8F推: 總之gene出問題就從protein同步再驗證06/29 00:02
2F推: 再釐清一下 所以你是將human tumor cell打到06/28 22:55
3F→: immunodeficient mice上,再做primary culture?06/28 22:56
8F推: DNA會影響RNA-seq的...先確定sample裡的DNA汙染程度06/28 23:23
9F→: 看個Fragment分析就知道了。也要問一下你的RNA-seq是否06/28 23:24
10F→: 可以排除DNA汙染 很久沒碰了 或許現在新技術可以無視XD06/28 23:25
1F推: 同樣的步驟Positive control 做得出來嗎?比如說raw+L08/12 00:53
2F→: PS08/12 00:53
8F推: 我沒碰過SW1353,至少要有TLRs之類的基因表現加LPS才08/14 13:04
9F→: 會有反應。另一條路是加東西誘導後直接用qPCR P看看有08/14 13:04
10F→: 沒有iNOS表現,如果跟control組差異不大就應該真的不08/14 13:04
11F→: 會有NO產生了,因為已經用兩種方法去驗證了。08/14 13:04
6F推: 冷凍切片可試試用-20度C的methanol fix11/14 23:33
7F→: 這樣可以不用打洞 不過還是得試試看11/14 23:34
1F→: sample是細胞?組織切片?11/05 01:58
2F→: 染膜蛋白也不一定要用到Triton 固定完直接blocking11/05 02:00
4F→: 一抗1hr+二抗0.5hr即可(室溫之下)11/05 02:03
13F推: PFA固定染螢光易有background 建議換固定buffer11/05 23:43
14F→: -20度C的methanol或是acetone都可以11/05 23:45
5F推:轉速大概1000~2000g就好 不然就加EDTA取plasma04/01 09:37
1F推:請問你的flow cytometry的機型是?10/23 20:45
4F推:如果用raw當model 測iNOS 測nitric oxide都可以09/23 20:49
5F推:之前用DAF-2DA這種probe 細胞染色、flow、ELISA都可以用09/23 20:56
2F推:HIF系列抗體推薦NOVUS07/19 20:21