Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

RNF40 (D2R2O) Rabbit mAb #12187

Ubiquitin Ligases  

No. Size Price
12187S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
12187 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human, Endogenous 130 Rabbit IgG
IP 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation,

Specificity / Sensitivity

RNF40 (D2R2O) Rabbit mAb recognizes endogenous levels of total RNF40 protein. This antibody does not cross-react with RNF20/BRE1A.

RNF40 (D2R2O) Rabbit mAb兔单抗能够检测内源性RNF40总蛋白水平。该抗体不会与RNF20/BRE1A 蛋白发生交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp537 of human RNF40 protein.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using RNF40 (D2R2O) Rabbit mAb.

Western blot方法检测不同细胞系的提取物,所用抗体为RNF40 (D2R2O) mAb兔单抗。

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human RNF20 (hRNF20-Myc/DDK; +) and Myc/DDK-tagged full-length human RNF40 (hRNF40-Myc/DDK; +), using RNF40 (D2R2O) Rabbit mAb (upper) or DYKDDDDK Tag Antibody #2368 (lower).

Western blot方法检测293T细胞提取物,分别用模拟物(-)或构建的带有Myc/DDK标签的全长人M RNF20 (hRNF20-Myc/DDK; +)以及带 Myc/DDK标签的全长人RNF40 (hRNF40-Myc/DDK; +),使用的抗体为 RNF40 (D2R2O) Rabbit mAb 兔单抗(上图) 以及DYKDDDDK Tag Antibody #2368 (下图)。



Immunoprecipitation of RNF40 from 293T cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or RNF40 (D2R2O) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using RNF40 (D2R2O) Rabbit mAb.

从293T细胞提取物中免疫沉淀RNF40,使用的抗体为Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane2)或RNF40 (D2R2O) Rabbit mAb (lane 3)兔单抗。Lane 1为10% input。 使用RNF40 (D2R2O) Rabbit mAb兔单抗进行Western blot检测。


In mammalian cells, the significance of histone H2B ubiquitination in chromatin epigenetics came from the identification of the budding yeast protein Bre1 (1,2). Together with the ubiquitin-conjugating enzyme Rad6, Bre1 serves as the E3 ligase in the monoubiquitination of the yeast histone H2B within transcribed regions of chromatin (1-3). Subsequently, the mammalian orthologs of yeast Bre1, RNF20 and RNF40, were identified (4,5). These two proteins form a tight heterodimer that acts as the major E3 ligase responsible for histone H2B monoubiquitination at Lys120 in mammalian cells, a modification linked to RNA Pol II-dependent transcription elongation in undamaged cells. Researchers have shown that DNA double-strand breaks (DSBs) are also capable of inducing monoubiquitination of H2B. This process depends upon the recruitment to DSB sites, as well as ATM-dependent phosphorylation of the RNF20-RNF40 heterodimer, thus highlighting a role for this E3 ligase in DSB repair pathways (6). Indeed, investigators have shown that loss of RNF20-RNF40 function promotes replication stress and chromosomal instability, which may constitute an early step in malignant transformation that precedes cell invasion (7).

在哺乳动物细胞中,histone H2B泛素化在染色质表观遗传学中的意义源于出芽酵母蛋白Bre1的确认(1,2)。Bre1蛋白与泛素偶联酶Rad6一起,可作为酵母histone H2B染色质转录区域的单泛素化中的E3连接酶(1-3)。随后,酵母Bre1蛋白的哺乳动物直系同源RNF20和RNF40蛋白被鉴定出来(4,5)。这两个蛋白质形成一个紧密的异源二聚体,可作为主要的E3连接酶使哺乳动物细胞中histone H2B蛋白Lys120位点单泛素化,这种修饰与非损伤细胞中RNA Pol II依赖的转录延伸相关。最近,研究显示DNA double-strand breaks (DSBs)也能够诱导H2B的单泛素化。与ATM依赖的RNF20-RNF40异源二聚体的磷酸化一样,这个过程取决于DSB位点上的募集,因此突出了这种E3连接酶在DSB修复通路中的作用 (6)。研究也的确显示了RNF20-RNF40功能的丧失会增加复制压力和染色体不稳定性,这可能形成了细胞侵袭前恶性转化的早期步骤(7)。

  1. Wood, A. et al. (2003) Mol Cell 11, 267-74.
  2. Hwang, W.W. et al. (2003) Mol Cell 11, 261-6.
  3. Kao, C.F. et al. (2004) Genes Dev 18, 184-95.
  4. Kim, J. et al. (2005) Mol Cell 20, 759-70.
  5. Zhu, B. et al. (2005) Mol Cell 20, 601-11.
  6. Moyal, L. et al. (2011) Mol Cell 41, 529-42.
  7. Chernikova, S.B. et al. (2012) Cancer Res, Epub ahead of print.

Application References

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