| 货号 | 5694S |
| 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 | RING1B (D22F2) XP® Rabbit mAb recognizes endogenous levels of total RING1B protein. |
| 使用方法 | WB(1:1000) IP (1:200) F (1:400) IF-IC (1:500) ChIP (1:50) |
| 供应商 | CST |
| 背景 | The polycomb group (PcG) proteins contribute 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 methyltransferase activity requires the Ezh2, Eed, and Suz12 subunits of the complex (5). Methylation of Lys27 facilitates the recruitment of the second complex, PRC1, which ubiquitinates histone H2A on Lys119 (6). PRC1 is composed of Bmi1 and RING1A (also RING1 or RNF1), both of which act to enhance the E3 ubiquitin ligase activity of an additional catalytic subunit RING1B (also RING2 or RNF2) (7). PcG proteins play an important role in the regulation of cell proliferation and senescence through repression of the p16 INK4A and p19 ARF genes and are required for maintenance of adult hematopoietic and neural stem cells, as well as embryonic stem cells (3,4,8-10). |
| 存放说明 | -20C |
| 计算分子量 | 41 |
Confocal immunofluorescent analysis of HeLa cells using RING1B (D22F2) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). 使用RING1B (D22F2) XP® Rabbit mAb (绿色)，共聚焦免疫荧光分析HeLa细胞。DY-554 phalloidin标记微丝蛋白(红色)。蓝色= DRAQ5® #4084 (DNA荧光染料)。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NCCIT cells and either 10 µl of RING1B (D22F2) XP® 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 RING1B (D22F2) XP® Rabbit mAb或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀实验。使用human HoxA1 intron 1 primers、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）。 | |
Western blot analysis of extracts from various cell lines using RING1B (D22F2) XP® Rabbit mAb. 使用RING1B (D22F2) XP® Rabbit mAb，免疫印迹(Western blot)分析不同细胞中RING1B (D22F2)的蛋白水平。 |
