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Antibodies

Histone H4K20me1 antibody (mAb)

Clone: 5E10-D8
Catalog No: 39727 Format: 100 µg $420 Buy Now
Catalog No: 39728 Format: 10 µg $95 Buy Now

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Antibody Type:Monoclonal
Isotype:IgG1k
Purification:Protein A Chromatography
Host:Mouse
Molecular Weight:8 kDa
Reactivity:Human, Other (Wide Range)

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated Immunofluorescence Validated Dot Blot Validated Immunocytochemistry Validated

for Histone H4 monomethyl Lys20 antibody (mAb) (Clone 5E10-D8)Application Notes


Validated Applications:
ChIP: 5 - 10 µg per ChIP
ChIP-Seq: 5 - 10 µg each
ICC/IF: 1 µg/ml dilution
WB: 0.5 - 2 µg/ml dilution

ChIP-Seq validation was performed by Active Motif's Epigenetics Services and the complete data set is available in the UCSC Genome Browser by clicking here.

Published Applications:
ChIP

for Histone H4 monomethyl Lys20 antibody (mAb) (Clone 5E10-D8)Immunogen

This Histone H4 monomethyl Lys20 was raised against a synthetic peptide containing monomethyl Lys20 of Histone H4.

for Histone H4 monomethyl Lys20 antibody (mAb) (Clone 5E10-D8)Buffer

Purified IgG in 70 mM Tris (pH 8), 105 mM NaCl, 31 mM glycine, 0.07 mM EDTA, 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

for Histone H4 monomethyl Lys20 antibody (mAb) (Clone 5E10-D8)References

 
Histone H4K20me1 antibody (mAb) tested by ChIP-Seq.

Histone H4K20me1 antibody (mAb) tested by ChIP-Seq.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 20 ug of chromatin from a human medulloblastoma cell line and 4 µg of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 12 million sequence tags were mapped to identify Histone H4K20me1 binding sites. The image shows binding across a region of chromosome 7. You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.

Histone H4K20me1 antibody (mAb) tested by immunofluorescence.

Histone H4 monomethyl Lys20 antibody tested by immunofluorescence.
Detection of Histone H4 monomethyl Lys20 by immunofluorescence. HeLa cells were stained with Histone H4 monomethyl Lys20 antibody at a dilution of 1 µg/ml. Top panel: Histone H4 monomethyl Lys20 antibody staining. Middle panel: DAPI. Bottom panel: merge.

Histone H4K20me1 antibody (mAb) tested by Western blot.

Histone H4 monomethyl Lys20 mAb (Clone 5E10-D8) tested by Western blot.
Detection of Histone H4 monomethyl Lys20 by Western blot. The analysis was performed using nuclear extract of HeLa cells probed with Histone H4 monomethyl Lys20 pAb (Clone 5E10-D8) at a dilution of 1 µg/ml.

Histone H4K20me1 antibody (mAb) tested by dot blot analysis.

Histone H4 monomethyl Lys20 mAb (Clone 5E10-D8) tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H4 monomethyl Lys20 mAb (Clone 5E10-D8) for monomethyl-lysine 20 of histone H4. Peptides corresponding to the immunogen and related peptides were spotted onto PVDF and probed with Histone H4 monomethyl Lys20 mAb (Clone 5E10-D8) at 1 ug/ml. The amount of peptide (picomoles) spotted is indicated next to each row.
Top panel: Lane 1: unmodified Lys79 peptide Lane 2: monomethyl Lys79 H4 Lane 3: dimethyl Lys79 H4 Lane 4: trimethyl Lys79 H4 Lane 5: monomethyl K20 H4 Lane 6: dimethyl K20 H4 Lane 7: trimethyl K20 H4 Bottom panel: Lane 1: monomethyl Lys31 H4 peptide Lane 2: dimethyl Lys31 H4 peptide Lane 3: trimethyl Lys31 H4 peptide Lane 4: monomethyl Lys44 H4 peptide Lane 5: dimethyl Lys44 H4 peptide Lane 6: trimethyl Lys44 H4 peptide.

for Histone H4 monomethyl Lys20 antibody (mAb) (Clone 5E10-D8)Background

Histone H4 is one of the core components of the nucleosome. The nucleosome is the smallest subunit of chromatin and consists of 147 base pairs of DNA wrapped around an octamer of core histone proteins (two each of Histone H2A, Histone H2B, Histone H3 and Histone H4). Chromatin is subject to a variety of chemical modifications, including post-translational modifications of the histone proteins and the methylation of cytosine residues in the DNA. Reported histone modifications include acetylation, methylation, phosphorylation, ubiquitylation, glycosylation, ADP-ribosylation, carbonylation and SUMOylation; these modifications play a major role in regulating gene expression.

Histone 4 lysine 20 (H4K20) can be mono-, di- or trimethylated by different histone methyltransferases such as NSD1 or ASH1. The methylation of this lysine is often associated with transcriptional repression. Monomethylation of histone H4 at lysine 20 is detected on the inactive X chromosome in female mammals and implicated in chromosome condensation.

for Histone H4 monomethyl Lys20 antibody (mAb) (Clone 5E10-D8)Positive Control

Active Motif's HeLa acid extract (Catalog No. 36200) can be used as a positive control for Histone H4 monomethyl Lys20 mAb (Clone 5E10-D8).

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with Histone H4K20me1 antibody (mAb).

Bridging Antibody to Improve ChIP, MeDIP and IP

Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use Histone H4K20me1 antibody (mAb).

Compatible Secondary Antibodies

Active Motif has developed a variety of high-quality secondary antibodies for most applications, with anti-rabbit and anti-mouse conjugates to a broad line of high-quality fluorescent molecules, as well as horse radish peroxidase (HRP). Visit our Secondary Antibody Conjugates page to find the perfect secondaries for immunofluorescence experiments with Histone H4K20me1 antibody (mAb).

for Histone H4 monomethyl Lys20 antibody (mAb) (Clone 5E10-D8)Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.

for Histone H4 monomethyl Lys20 antibody (mAb) (Clone 5E10-D8)Guarantee

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer;
  • WB = Western Blotting
 

Technical Data Sheet

Histone H4K20me1 antibody (mAb)
 

Data Thumbnails

Histone H4K20me1 antibody (mAb) tested by ChIP-Seq.

ChIP-Seq of Histone H4K20me1 mAb.
(Click image to enlarge and see details.)

Histone H4K20me1 antibody (mAb) tested by immunofluorescence.

Immunofluorescence stain of Histone H4K20me1 mAb.
(Click image to enlarge and see details.)

Histone H4K20me1 antibody (mAb) tested by Western blot.

Western blot of Histone H4K20me1 mAb.
(Click image to enlarge and see details.)

Histone H4K20me1 antibody (mAb) tested by dot blot analysis.

Dot blot of Histone H4K20me1 mAb.
(Click image to enlarge and see details.)