| 货号 | 5856S |
| 反应种属 | Human/Mouse/Rat/Monkey |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IF-IC/ChIP |
| 目标/特异性 | Bmi1 (D42B3) Rabbit mAb recognizes endogenous levels of total Bmi1 protein. |
| 使用方法 | WB(1:1000) IP (1:100) IF-IC (1:100) ChIP (1:50) |
| 供应商 | CST |
| 背景 | The polycomb group (PcG) of proteins contributes to the maintenance of cell identity, stem cell self-renewal, cell cycle regulation, and oncogenesis by maintaining the silenced state of genes that promote cell lineage specification, cell death, and cell-cycle arrest (1-4). PcG proteins exist in two complexes that cooperate to maintain long-term gene silencing through epigenetic chromatin modifications. The first complex, EED-EZH2, is recruited to genes by DNA-binding transcription factors and methylates histone H3 on Lys27. This histone methyl-transferase activity requires the Ezh2, Eed, and Suz12 subunits of the complex (5). Histone H3 methylation at Lys27 facilitates the recruitment of the second complex, PRC1, which ubiquitinylates histone H2A on Lys119 (6). Bmi1 is a component of the PRC1 complex, which together with Ring1 strongly enhances the E3 ubiquitin ligase activity of the Ring2 catalytic subunit (7). Bmi1 plays an important role in the regulation of cell proliferation and senescence through repression of the p16 INK4A and p19 ARF genes and is required for maintenance of adult hematopoietic and neural stem cells (3,4,8-10). |
| 存放说明 | -20C |
| 计算分子量 | 41, 43 |
Confocal immunofluorescent analysis of C2C12 cells using Bmi1 (D42B3) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). 使用Bmi1 (D42B3) Rabbit mAb 兔单抗(绿色)标记，共聚焦免疫荧光分析C2C12细胞。DY-554 phalloidin标记微丝蛋白(红色)。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NCCIT cells and either 10 µl of Bmi1 (D42B3) Rabbit mAb or 2 µl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human HoxA1 intron 1 primers, SimpleChIP® Human HoxA2 Promoter Primers #5517, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin (equivalent to one). 使用SimpleChIP®Enzymatic Chromatin IP Kit (Magnetic Beads) #9003，用4 x 106 NCCIT细胞的交联染色质以及10 µl Bmi1 (D42B3) Rabbit mAb或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀实验。使用SimpleChIP®Human HoxA1 Intron 1 Primers #7707、SimpleChIP® Human HoxA2 Promoter Primers #5517和SimpleChIP® Human α Satellite Repeat Primers #4486，浓缩的DNA通过real-time PCR定量。在每个样品中免疫沉淀DNA的数量被当做一个相对于总input chromatin的数量的信号(相当于一个)。 | |
Western blot analysis of extracts from various cell lines using Bmi1 (D42B3) Rabbit mAb. 使用Bmi1 (D42B3) Rabbit mAb兔单抗，免疫印迹(Western blot)分析不同细胞中Bmi1蛋白水平。 |
