Our cell-cycle reporter vectors allow for real-time monitoring of cell-cycle progression in living cells without fixation. They even let you visualize cell shape.
Our cell-cycle reporter vectors allow for real-time monitoring of cell-cycle progression in living cells without fixation. They even let you visualize cell shape.
Fucci probes
Cell-cycle reporter vectors deliver fluorescent, ubiquitination-based cell-cycle indicators (Fucci) that allow you to identify cells in various phases of the cell cycle. These Fucci probes contain Cdt1 or Geminin, proteins whose levels fluctuate differentially throughout the cell cycle: Cdt1 levels peak in G1 phase; as cells transition into S phase, Cdt1 levels fall and Geminin levels rise, remaining high until the cells are back in G1. Cells control Cdt1 and Geminin levels post-translationally, using ubiquitination to target the unwanted proteins for proteasomal degradation. To allow Geminin and Cdt1 to be easily visualized, each is expressed with a red or cyan Living Colors fluorescent tag. These probes allow precise, visual evaluation of the cell-cycle phase.
Monitor cell cycle and cell shape in real time
Our Fucci cell-cycle reporters let you label just the nucleus, or both the nucleus and cytoplasm, allowing visualization of cell shape. Although most Fucci probes label only cell nuclei, we offer a truncated version of hGeminin that is able to migrate to the cytoplasm between S and M phases, enabling visualization of cell morphology.
A variety of Fucci probes and delivery options
We offer retrovirus and plasmid-based vectors for the delivery of a variety of Fucci probes. Each probe consists of a red or cyan fluorescent tag, and a deletion mutant of either hCdt1 or hGeminin. Our retroviral vectors can either be transfected as plasmids or transduced as retrovirus in a wide variety of mammalian cells. Once integrated into the host genome, these vectors provide stable, heritable expression of the probes.
The plasmid-based vector, pTRE-CellCycle, allows tightly controlled, tetracycline (Tet)-inducible expression of two Fucci probes from a bidirectional promoter that can be induced using any Tet-On or Tet-Off technology. Expression of these probes causes cell nuclei to turn red during G1 phase and cyan during S, G2, and M phases, allowing complete visual tracking of the cell cycle.
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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
