Histones are highly conserved proteins that serve as the structural scaffold for the organization of nuclear DNA into chromatin. The four core histones, H2A, H2B, H3, and H4, assemble into an octamer (2 molecules of each). Histones are modified post-translationally by the actions of enzymes in both the nucleus and cytoplasm. These modifications, which include acetylation, phosphorylation, methylation, and ubiquitin-ation, occur predominantly on the N-terminal and C-terminal tails that extend beyond the nucleosome core particle. Trimethylation of histone H4 on lys20 (H4-K20me3) selectively marks constitutive pericentromeric hetero-chromatin and imprinting control regions, while mono- and dimethylation are broadly distributed but principally enriched in euchromatin regions. Although the biological function of H4K20me remains poorly understood, studies suggest that H4-K20me2 is involved in the guidance of DNA repair proteins to DNA strand breaks, whereas H4K20me1 is associated with chromatin condensation processes. PR-Set7 (also known as SET8) specifically catalyzes monomethylation of H4K20me1.

Recognizes monomethyl-histone H4 (Lys20), Mr ~10kDa.

The immunizing sequence is conserved from Tetrahymena to human, so broad species cross-reactivity is expected.

KLH-conjugated, synthetic peptide containing the sequence …HR[mmK]V…, in which mmK corresponds to monomethyl-lysine at residue 20 of human histone H4.

Immunoblot Analysis:
A 1:4000 dilution of a previous lot detected methylated histone H3 in acid extracted proteins from HeLa cells. The antibody did not detect unmethylated recombinant Histone H3.

Multiplexing:
Beadlyte Histone-Peptide Specificity Assay: 1:2,000-1:30,000 dilutions of a previous lot were incubated with histone H4 peptides containing various modifications conjugated to Luminex microspheres. Specificity to monomethyl-histone H4 (Lys20) was demonstrated (Figure C).

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Histones

Use Anti-monomethyl-Histone H4 (Lys20) Antibody, clone NL314 (rabbit monoclonal antibody) validated in WB, Mplex to detect monomethyl-Histone H4 (Lys20) also known as H4K20me1, Histone H4 (mono methyl K20).

Routinely evaluated by immunoblot analysis.
Immunoblot Analysis:
A 1:1000-1:2000 dilution of this lot detected monomethyl histone H4 (Lys20) in HeLa acid extracts. (Figure A).

10kDa

Cultured supernantant in 0.05% sodium azide

Stable for 1 year at -20°C from date of receipt.
For maximum recovery of product, centrifuge the vial prior to removing the cap.

Control
HeLa acid extracts

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