2017 Guide of H4k20 methylation antibodies

 

1. Background information of H4K20me

Methylation of H4K20 is playing an important role with several nuclear processes, including transcriptional silencing, DNA damage response, DNA replication, and maintenance of genome integrity. H4K20 methylation is primarily mediated by two KMTs in metazoans, one is SET8 (also known as PR-SET7 and KMT5A) and another is SUV4-20, which possesses two homologs H1 and H2 in mammals. SET8 is an H4K20 monomethyltransferase, and SUV4-20H1/2 was first reported to function as H4K20 di- and trimethyltransferases that associate with pericentric heterochromatin according to biochemical and structural studies. Further studies have found SET8 and SUV4-20 has a more nuanced relationship in establishing H4K20methylation states. In Drosophila, a decrease in H4K20me2/3 coupled with an accumulation of H4K20me1 is caused by RNA interference (RNAi)-mediated knockdown and null mutations of SUV4-20, whereas a marked increase in unmodified H4K20 is affected by RNAi knockdown of SET8. And a mouse study also proved the SUV4-20 results. Together, the model of H4K20 methylation is including  SET8 is mainly responsible for generating H4K20me1 and SUV4-20 then methylates H4K20me1 to increase H4K20me2 and H4K20me3 level.

2. Three types of H4k20 methylation: H4k20me1, H4k20me2, H4k20me3

SUV4-20H1/2 was first reported to function as H4K20me2 and H4K20me3 that associate with pericentric heterochromatin and these KMTs predominantly using H4K20me1 as a substrate to produce H4K20me2 by more recent functional and structural studies. Regulatory covalent modifications of the SUV4-20 KMTs or their association with heterochromatic factors may change their product specificity toward H4K20me3 are two possible ways of H4K20me3 regulation. Essential for deposition of H4K20me3 in heterochromatin by SUV4-20 in Drosophila is heterochromatin protein 1 (HP1) suggesting that H4K20 trimethyltransferase activity may stimulate by HP1. Alternatively, the human genome encodes over 50 SET proteins, but less than half of them have been biochemically characterized to date, leaving the possibility that additional H4K20-specific KMTs exist. The mechanism by the H4K20me3 modification is established and propagated in pericentric heterochromatin needs further studies to figure out.

 

Check 2017 ChIP Validated Histone H4k20me antibodies. Epicypher validated Abcam OEM supplier: 

More details about the supplier

Point 1
Validated Original manufacturer.

ABclonal Inc. is one of the original manufactures which is famous for its Epigenetics antibodies such as H3K4me antibody and H3K9me antibody. It is the long-term OEM supplier for many top10 antibody companies and we also visited their factory for double confirmation.

Point 2
Lot number management

Lot number is unique for each batch of antibody and it can be taken as identity number of antibody.  ABclonal has effective batch management and it helps you maximize the reproducibility. We will share everything about the antibodies basing on Lot number.

Point 3
Antibody validation for each batch.

ABclonal is performing strict antibody validation for each batch. WB, IHC, IF, DB and ChIP are performed to make sure each Histone antibody has strong specificity and no cross-reactivity for multiple applications. It is the only one of 52 Histone antibody suppliers which passed ChIP test by EpiCypher in 2017. (Paper to be published)


3. Relating protein study of H4K20me:
The H34K20me protein study has two parts, including writers and readers.
PR-SET7/ SETD8, SUV420H1&2 and NSD1/2 are writers
L3MBTL1, SFMBT, MBTD1 and 53BP1 are readers.

Writers Find antibodies Introduction Reference
PR-SET7/ SETD8 SETD8 antibody PR-SET7/SETD8 is able to monomethylate H4K20me1. It controls both H4K16Ac and H4K20me3 and in doing so. It also regulates Pol II pausing dynamics. PR-SET7-mediated H4K20me1 is necessary for the recruitment of the MSL complex, release of Pol II into active elongation, and subsequent H4K16Ac Kapoor-Vazirani, Priya; Vertino, Paula M. Journal of Biological Chemistry. 3/14/2014, Vol. 289 Issue 11, p7425-7437. 13p. DOI: 10.1074/jbc.M113.520783. , Database: Academic Search Complete
SUV420H1&2   SUV420H1 and SUV420H2 are two strongly homologous enzymes that methylate H4K20. H4K20 is a mark that has been influenced in transcriptional regulation. Wu, H; Siarheyeva, A; Zeng, H; Lam, R; Dong, AP; Wu, XH; Li, YJ; Schapira, M; Vedadi, M; Min, J. FEBS LETTERS; NOV 29, 2013; 587; 23; p3859-p3868, Database: Science Citation Index
NSD1/2 NSD1 antibody

 

NSD2 antibody
Initially, NSD1 was reported to methylate H3K36 and H4K20. Then contradicting data concerning NSD1’s specificity were published: a study documented that NSD1 depletion did not change the status of H4K20 methylation, while another reported that reduced expression of NSD1 led to the reduction of H4K20 methylation. Similarly, research data from the H4K20 methylation activity of NSD2 were also controversial. Kudithipudi, Srikanth; Lungu, Cristiana; Rathert, Philipp; Happel, Nicole; Jeltsch, Albert. In Chemistry & Biology. 20 February 2014 21(2):226-237 Language: English. DOI: 10.1016/j.chembiol.2013.10.016, Database: ScienceDirect
Readers Find antibodies Introduction Reference
L3MBTL1 L3MBTL1 antibody Lethal 3 malignant brain tumor 1 (L3MBTL1), a homolog of the Drosophila polycomb tumor suppressor l (3)mbt. L3MBTL1 contains three tandem MBT repeats (3xMBT). And 3xMBT are critical for transcriptional repression. The wide range of in vitro binding, the chromatin association of L3MBTL1 mirrors the progressive accumulation of H4K20me1 during the cell cycle. Kalakonda, N; Fischle, W; Boccuni, P; Gurvich, N; Hoya-Arias, R; Zhao, X; Miyata, Y; MacGrogan, D; Zhang, J; Sims, J K; Rice, J C; Nimer, S D. Oncogene. 7/17/2008, Vol. 27 Issue 31, p4293-4304. 12p. 2 Black and White Photographs, 1 Chart, 4 Graphs. DOI: 10.1038/onc.2008.67. , Database: Academic Search Complete
SFMBT  According to research, polycom group was repressed by Mammalian SFMBTs. SFMBT2 has a transcriptional repression activity on HOXB13 gene expression by using DU145 prostate cancer cells as a model. At the HOXB13 gene promoter, occupancy of SFMBT2 coincided with enrichment of H3K9me2/3 and H4K20me2/3 as well as H3K27me3. Lee, K; Na, W; Maeng, JH; Wu, H; Ju, BG. JOURNAL OF BIOSCIENCES; MAR, 2013; 38; 1; p105-p112, Database: Science Citation Index
MBTD1 and 53BP1 53BP1 (TP53BP1)antibody MBTD1 is a stable subunit of the TIP60/NuA4 complex. MBTD1 as part of TIP60 regulates transcription and repair of DNA breaks. MBTD1 competes with 53BP1 for binding to the H4K20me mark. Jacquet, Karine; Fradet-Turcotte, Amélie; Avvakumov, Nikita; Lambert, Jean-Philippe; Roques, Céline; Pandita, Raj K.; Paquet, Eric; Herst, Pauline; Gingras, Anne-Claude; Pandita, Tej K.; Legube, Gaëlle; Doyon, Yannick; Durocher, Daniel; Côté, Jacques. In Molecular Cell. 5 May 2016 62(3):409-421 Language: English. DOI: 10.1016/j.molcel.2016.03.031, Database: ScienceDirect

Reference

Del Rizzo, Paul A.; Trievel, Raymond C.. In Methylation: A Multifaceted Modification – looking at transcription and beyond, BBA – Gene Regulatory Mechanisms. December 2014 1839(12):1404-1415 Language: English. DOI: 10.1016/j.bbagrm.2014.06.008, Database: ScienceDirect