Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

RING1B (D22F2) XP® Rabbit mAb #5694

chip-seq   polycomb   PRC   PRC1   ring1  

No. Size Price
5694S 100 µl ( 10 western blots ) ¥3,580.00 现货查询 购买询价
5694T 20 µl ( 2 western blots ) ¥1,400.00 现货查询 购买询价
5694 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 41 Rabbit IgG
IP 1:200
F 1:400
IF-IC 1:500
ChIP 1:50
ChIP-seq 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP, ChIP-seq=Chromatin IP-seq,

Specificity / Sensitivity

RING1B (D22F2) XP® Rabbit mAb recognizes endogenous levels of total RING1B protein.

RING1B (D22F2) XP® Rabbit mAb兔单抗能够检测内源性RING1B总蛋白水平。

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the full-length human RING1B protein.




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 IP

Chromatin IP

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 Blotting

Western Blotting

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。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells using RING1B (D22F2) XP® Rabbit mAb (blue) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Chromatin IP-seq

Chromatin IP-seq

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 10 µl of Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb #9733, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared from 5ng enriched ChIP DNA for RING1B ChIP-seq and 50ng enriched ChIP DNA for H3K27me3 ChIP-seq using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina®, and sequenced on the Illumina NextSeq. RING1B and H3K27me3 are known to associate with each other on chromatin. The figure shows binding of both RING1B and H3K27me3 across HOXD genes, which are known target genes of RING1B (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.


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).

polycomb group (PcG)蛋白有助于细胞身份的维持、干细胞自我更新、细胞周期调节和肿瘤形成,这些功能通过维持一些基因的沉默,而这些基因是促进细胞系特异性、细胞死亡和细胞周期停滞(1-4)。PcG蛋白存在两个复合物,它们配合通过表观遗传染色质修饰去维持长期的基因沉默。通过DNA结合转录因子和组蛋白H3在Lys27位点的甲基化,第一个复合物EED-EZH2被招募到基因上。这个组蛋白甲基转移酶活性需要该复合物的Ezh2、Eed和Suz12亚基(5)。Lys27位点的甲基化促进第二个复合物PRC1的招募,其能使组蛋白H2A在Lys119位点泛素化(6)。PRC1是由Bmi1和RING1A (RING1或RNF1)组成,它们可提高一个额外的催化亚单位RING1B (RING2或RNF2)的E3泛素连接酶活性(7)。PcG蛋白在调节细胞增殖和衰老中起到重要作用,这主要通过p16 INK4A和p19 ARF基因的抑制,以及像胚胎干细胞一样对于成人造血和神经干细胞的维持是需要的(3,4,8-10)。

  1. Boyer, L.A. et al. (2006) Nature 441, 349-53.
  2. Lee, T.I. et al. (2006) Cell 125, 301-13.
  3. Park, I.K. et al. (2003) Nature 423, 302-5.
  4. Molofsky, A.V. et al. (2003) Nature 425, 962-7.
  5. Cao, R. and Zhang, Y. (2004) Mol Cell 15, 57-67.
  6. Wang, H. et al. (2004) Nature 431, 873-8.
  7. Cao, R. et al. (2005) Mol Cell 20, 845-54.
  8. Molofsky, A.V. et al. (2005) Genes Dev 19, 1432-7.
  9. Jacobs, J.J. et al. (1999) Nature 397, 164-8.
  10. Jacobs, J.J. et al. (1999) Genes Dev 13, 2678-90.

Application References

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