Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

RNF20 (D6E10) XP® Rabbit mAb #11974

No. Size Price
11974S 100 µl ( 10 western blots ) ¥3,750.00 现货查询 购买询价 防伪查询
11974 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 120 Rabbit IgG
IP 1:200
ChIP 1:50

Species cross-reactivity is determined by western blot.

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


Species predicted to react based on 100% sequence homology: Hamster, Dog, Pig, Guinea Pig, Horse,

Specificity / Sensitivity

RNF20 (D6E10) XP® Rabbit mAb recognizes endogenous levels of total RNF20 protein. This antibody recognizes a second band at 80 kDa in mouse lysates, corresponding to mouse RNF20 isoform 2. This antibody does not cross-react with RNF40 protein.

RNF20 (D6E10) XP® Rabbit mAb兔单抗能够检测内源性RNF20总蛋白。该抗体还能够识别另外一个小鼠裂解物中80kDa的条带,对应的是小鼠RNF20异构体2。该抗体不会与RNF40蛋白产生交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly516 of human RNF20 protein.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using RNF20 (D6E10) XP® Rabbit mAb.

Western blot方法检测不同细胞系的提取物,所用抗体为RNF20 (D6E10) XP® Rabbit mAb兔单抗。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 μl of RNF20 (D6E10) 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 SimpleChIP® Human γ-Actin Promoter Primers #5037, SimpleChIP® Human GAPDH Promoter Primers #4471, 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, which is equivalent to one.

使用4 x 106 HeLa细胞中交联过的染色质以及10 µl RNF20 (D6E10) XP® Rabbit mAb 或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀,所用试剂盒为SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003。富集到的DNA经过实时PCR定量,所使用产品为SimpleChIP® Human γ-Actin Promoter Primers #5037、 SimpleChIP® Human GAPDH Promoter Primers #4471以及 SimpleChIP® Human α Satellite Repeat Primers #4486。每个样品中免疫沉淀得到的DNA量由与input chromatin( 相当于1)的相对信号值来表示。


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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For Research Use Only. Not For Use In Diagnostic Procedures.

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