The Mir-X inducible miRNA red and green systems exploit key elements of our Tet-On Advanced Inducible Expression System to provide controlled co-expression of your selected microRNA and a bright red or green fluorescent protein marker.
The Mir-X inducible miRNA red and green systems exploit key elements of our Tet-On Advanced Inducible Expression System to provide controlled co-expression of your selected microRNA and a bright red or green fluorescent protein marker.
How it works
Your microRNA sequence is embedded in the 3’ UTR of a marker protein mRNA, which is expressed from an inducible miRNA expression vector (pmRi-mCherry or pmRi-ZsGreen1). When cells expressing the Tet-On Advanced transactivator are transfected with the pmRi-mCherry or pmRi-ZsGreen1 vector, high levels of your inducible microRNA and either the mCherry or ZsGreen1 proteins are expressed in response to doxycycline (Dox). Once Dox is added to the cell culture medium, miRNA induction is very rapid and effective.
We used modified Metridia-secreted luciferase genes containing miRNA target sequences (RNAiMonitor) to measure the effects of miR-1 and miR-9 miRNA induction in MCF7 Tet-On Advanced cells. Inducing miR-1 and miR-9 expression in these cells with Dox resulted in >90% knockdown of luciferase activity compared to controls.
Red and green complete systems
The complete Mir-X inducible miRNA systems provide the pTet-On Advanced Vector for expressing the Dox-responsive transactivator; an inducible miRNA expression vector (either pmRi-mCherry or pmRi-ZsGreen1, also available separately); two linear selectable markers (hygromycin and puromycin) for cotransfecting with the pmRi vectors in order to establish stable cell lines; and tetracycline-free Tet System Approved FBS (serum) for ensuring the full activity of Tet-based expression systems.
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请问各位大神，需要做人巨噬细胞的F4/80,CD11b，CD163标记物，用流式，抗体哪家比较好呢，请推荐，谢谢！
比如，你使用了小鼠IgG1, PE 标记的特异性抗体，那就选用小鼠IgG1， PE标记的非特异性抗体。
下面这个图，是一个比较典型的使用了同型对照的结果图向左转|向右转灰色的曲线是未染色的样本，黄色是同型对照，红色是特异性抗体染色。由于细胞对抗体的非特异性结合，一般同型对照都会比未染色样本的荧光信号要强。所以设置阴性阈值的时候，要根据同型对照，而不是未染色的细胞。
请问一下做流式需要的抗体那种比较好，有人说Biolegend，eBioscience，santa，BD。求各位大神指教
如果一个抗体可以用来“做western blot，免疫荧光，免疫共沉淀,ELISA”，我估计着是HRP或碱性磷酸酶或生物素之类标记的，然后需要再加荧光标记的二抗，这类抗体也是不能做流式用的。
我只用过流式的抗体来做免疫荧光（直接染色），效果还不错，但是反过来就没试过。
楼上两位还是要仔细看下产品说明书
