Histone H4R3me2a (asymmetric) antibody (pAb)

Catalog No: 39705 Format: 100 µl $435 Buy
Catalog No: 39006 Format: 50 µl $250 Buy
Catalog No: 39706 Format: 10 µl $105 Buy

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Antibody Type:
Polyclonal
Isotype:
IgG
Purification:
Protein A Chromatography
Host:
Rabbit
Molecular Weight:
8 kDa
Reactivity:
Human, Other (Wide Range)

Applications

ChIP Validated Western Blot Validated Immunofluorescence Validated Dot Blot Validated

Application Notes


Validated Applications:
WB: 1:500 dilution

Published Applications:
ChIP
WB
IF
See references for more information. Individual optimization may be required.

Immunogen

This Histone H4 dimethyl Arg3, asymmetric antibody was raised against a synthetic peptide containing dimethyl-Arg3 (asymmetric) of human histone H4.

Buffer

Purified rabbit IgG in PBS with 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

References

 
Histone H4R3me2a (asymmetric) antibody (pAb) tested by Western blot.

Histone H4R3me2a (asymmetric) antibody (pAb)tested by Western blot.
Nuclear extract of HeLa cells (20 µg) tested by Western blot using Histone H4R3me2a (asymmetric) antibody (pAb) at a dilution of 1:500.

Histone H4R3me2a (asymmetric) antibody (pAb) tested by dot blot analysis.

Histone H4R3me2a (asymmetric) antibody (pAb)tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H4 dimethyl Arg3 asymmetric antibody for dimethyl-arginine 3 of histone H4. Peptides corresponding to the immunogen and related peptides were spotted onto PVDF and probed with Histone H4 dimethyl Arg3 asymmetric antibody at 1:2,500. The amount of peptide (picomoles) spotted is indicated next to each row.

Lane 1: unmodified H4 peptide aa 1-10.
Lane 2: H4 monomethyl-Arg3 peptide.
Lane 3: dimethyl-Arg3(asymmetric) H4 peptide.
Lane 4: dimethyl-Arg3 (symmetric) H4 peptide.

Histone H4R3me2a antibody (pAb) specificity tested by peptide array analysis.

Histone H4R3me2a (asymmetric) antibody (pAb) specificity tested by peptide array analysis.
Peptide array analysis was used to confirm the specificity of this antibody for its intended modification. Histone H4R3me2a antibody was applied at a dilution of 1:20,000 to Active Motif's MODified™ Histone Peptide Array (Catalog No. 13001). The arrays were scanned with ArrayAnalysis Software 16 and the results plotted. Specificity data is shown for the most reactive peptides and those related to the immunogen.

Background

Histone H4 is one of the core components of the nucleosome, the basic building block of chromatin. Histones are 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. The methylation of histones can occur on two different residues: arginine or lysine.

Asymmetric dimethylation of histone H4 at arginine 3 (Arg3), catalyzed by the PRMT methyltransferases, is associated with transcriptional activation by nuclear hormone receptors. Arg3 methylation has been reported to facilitate the subsequent acetylation of histone H4 by the acetyltransferase p300. Histone H4 dimethyl Arg3 asymmetric methylation (H4R2me2a) is catalyzed by PRMT1 and PRMT6.

Positive Control

Active Motif's HeLa acid extract (Catalog No. 36200) can be used as a positive control for Histone H4 dimethyl Arg3, asymmetric antibody.

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 H4R3me2a (asymmetric) antibody (pAb).

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 H4R3me2a (asymmetric) antibody (pAb).

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.

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