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Key Documents

07-1570

Sigma-Aldrich

Anti-monomethyl Histone H4 (Lys20) Antibody

from rabbit, purified by affinity chromatography

Synonym(s):

H4K20me1, Histone H4 (mono methyl K20), H4 histone family, member A, histone 1, H4a, histone cluster 1, H4a

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41

biological source

rabbit

Quality Level

antibody form

affinity isolated antibody

antibody product type

primary antibodies

clone

polyclonal

purified by

affinity chromatography

species reactivity

human

technique(s)

ChIP: suitable
dot blot: suitable
inhibition assay: suitable (peptide)
western blot: suitable

NCBI accession no.

UniProt accession no.

shipped in

dry ice

target post-translational modification

monomethylation (Lys20)

Gene Information

human ... H4C1(8359)

General description

Histones are highly conserved proteins that serve as the structural scaffold for the organization of nuclear DNA into chromatin. Histones modifications regulate DNA transcription, repair, recombination, and replication. The most commonly studied modifications are acetylation, phosphorylation, methylation, and ubiquitination. These modifications can alter local chromatin architecture, or recruit trans-acting factors that recognize specific histone modifications (the "histone code" hypothesis). Methylation of histone H4 at lysine 20 has been implicated in transcriptional activation, gene silencing, heterochromatin formation, mitosis, and DNA repair. Histone H4 is progressively methylated at Lys20 during the G(2), M, and G(1) phases of the cell cycle, and methylation of H4Lys20 is a marker for heterochromatin. In mammals, the female X chromosome is coated by the Xist RNA, which is critical for silencing, and is marked mainly by methylation of H3Lys9, H3Lys27 and H4Lys20.

Specificity

Based on sequence homology, broad species cross-reactivity is expected
Recognizes monomethylated Histone H4
(Lys20), Mr 11 kDa.

Application

Anti-monomethyl Histone H4 (Lys20) Antibody is a rabbit polyclonal antibody for detection of monomethyl Histone H4 (Lys20) also known as H4K20me1, Histone H4 (mono methyl K20), H4 histone family member A & has been validated in WB.
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (2 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 4 μg of either a negative control antibody or Anti-Monomethyl-Histone H4
(Lys20) antibody and the Magna ChIP A Kit (Cat. #17-610). Successful immunoprecipitation of monomethyl-histone H4 (Lys20)-associated DNA fragments was verified by qPCR using GAPDH coding region ChIP Primers versus Control Primers corresponding to the GAPDH promoter (Please see figures). Data is presented as percent input of each IP sample relative to input chromatin, with immunoprecipitated DNA from negative control antibody shown as (-) and monomethyl-histone H4 (Lys20) shown as (+).
Please refer to the EZ-Magna ChIP A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.

Dot Blot Analysis :
Absurance Histone H3 Antibody Specificity Array (Cat. No. 16-667) and Absurance Histone H2A, H2B, H4 Antibody Specificity Array (Cat. No. 16-665), which contain histone peptides with various modifications were probed with Cat. No. 07-1570 Anti-monomethyl Histone H4 (Lys20) at 1:1000 dilution. Proteins were visualized using a Donkey anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system.

Peptide Blocking Assay:
40 μg of histone H4 peptide containing monomethyl lysine 20 abolished detection of histone H4 by anti-monomethyl-Histone H4 (Lys20) in immunoblot analysis of acid extracts of HeLa cells

Quality

Routinely evaluated by western blot analysis

Target description

~11 kDa

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Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

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A trans-tail histone code defined by monomethylated H4 Lys-20 and H3 Lys-9 demarcates distinct regions of silent chromatin.
Sims, Jennifer K, et al.
The Journal of Biological Chemistry, 281, 12760-12766 (2006)
James J Pesavento et al.
Molecular and cellular biology, 28(1), 468-486 (2007-10-31)
Methylation of histone H4 at lysine 20 (K20) has been implicated in transcriptional activation, gene silencing, heterochromatin formation, mitosis, and DNA repair. However, little is known about how this modification is regulated or how it contributes to these diverse processes.
Radiation-induced alterations of histone post-translational modification levels in lymphoblastoid cell lines.
Maroschik, B; Gurtler, A; Kramer, A; Ro?ler, U; Gomolka, M; Hornhardt, S; Mortl, S; Friedl, AA
Radiation Oncology (London, England) null
H Nakshatri et al.
Cell death & disease, 6, e1608-e1608 (2015-01-23)
The transcription factor nuclear factor-kappaB (NF-κB) is constitutively active in several cancers and is a target of therapeutic development. We recently developed dimethylaminoparthenolide (DMAPT), a clinical grade water-soluble analog of parthenolide, as a potent inhibitor of NF-κB and demonstrated in

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