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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.
~11 kDa
Immunogen
Epitope: Monomethyl Lys20
KLH-conjugated, linear synthetic peptide corresponding to amino acids 15-24
(AKRHR[me1K]VLRD-C) of human Histone H4.
Application
Anti-monomethyl Histone H4 (Lys20) Antibody, Trial Size 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 refer to the EZ-Magna ChIP
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-S 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.
Western Blot Analysis:
0.1 μg/mL of proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures)
Peptide Blocking Assay:
40 μg of histone H4 peptide containing monomethyl Lys20 abolished detection of histone H4 by anti-monomethyl-Histone H4 (Lys20) in immunoblot analysis of acid extracts of HeLa cells (Please see figures).
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Chromatin Biology
Histones
Biochem/physiol Actions
Based on sequence homology, broad species cross-reactivity is expected
Recognizes monomethylated Histone H4
(Lys20), Mr 11 kDa.
Physical form
Affinity Chromatography
Purified from serum by affinity chromatography. Supplied at 1 mg/ml in 0.1M Tris-Glycine (pH7.4), 15mM NaCl, 0.05% NaN3.
Preparation Note
Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Analysis Note
Control
HeLa cell acid extracts.
Routinely evaluated by western blot analysis
Legal Information
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
| biological source | rabbit |
| Quality Level | 100 |
| antibody form | affinity isolated antibody |
| antibody product type | primary antibodies |
| clone | polyclonal |
| purified by | affinity chromatography |
| species reactivity | human |
| manufacturer/tradename | Upstate® |
| technique(s) | ChIP: suitable,dot blot: suitable,inhibition assay: suitable (peptide),western blot: suitable |
| NCBI accession no. | NM_003538.3 |
| UniProt accession no. | P62805 |
| shipped in | wet ice |
| target post-translational modification | monomethylation (Lys20) |
| Gene Information | human ... H4C1(8359) |
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