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Antibodies

AbFlex® Histone H3K4me3 antibody (rAb)

Catalog No: 91263 Format: 100 µg $375 Buy Now
Catalog No: 91264 Format: 10 µg $75 Buy Now

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Antibody Type:Recombinant
Isotype:IgG2a
Purification:Protein A Chromatography
Host:Mouse
Molecular Weight:17 kDa
Reactivity:Human, Other (Wide Range)

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated Dot Blot Validated

for AbFlex Histone H3K4me3 antibody (rAb)Application Notes


Validated Applications:
ChIP-Seq: 4 µg/ml
WB: 1 - 2 µg/ml
Dot Blot: 1 µg/ml

Note: many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt / Sonication Protocol when preparing nuclear extracts for Western Blot.

for AbFlex Histone H3K4me3 antibody (rAb)Immunogen

This antibody was raised against a synthetic peptide containing trimethyl Lys4 of human histone H3.

for AbFlex Histone H3K4me3 antibody (rAb)Buffer

Purified IgG in 140 mM Hepes, pH 7.5, 70 mM NaCl, 32 mM NaOAc, 0.035% sodium azide, 30% glycerol.

View our Guide to Histone Modifications and Biological Function.

 
AbFlex<sup>®</sup> Histone H3K4me3 antibody (rAb) tested by ChIP-Seq.

AbFlex® Histone H3K4me3 recombinant antibody (rAb) tested by ChIP-Seq
Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 30 µg of human acute myelocytic leukemia chromatin and 4 µg of AbFlex Histone H3K4me3 antibody. ChIP DNA was sequenced on the Illumina NextSeq and 8 million sequence tags were mapped to identify Histone H3K4me3 binding sites on chromosome 7. You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.

AbFlex<sup>®</sup> Histone H3K4me3 antibody (rAb) tested by Western blot.

AbFlex® Histone H3K4me3 antibody (rAb) tested by Western blot.
20 µg of HeLa cell extract was run on SDS-PAGE and probed with AbFlex Histone H3K4me3 antibody at 2 µg/ml

AbFlex<sup>®</sup> Histone H3K4me3 antibody (rAb) tested by dot blot analysis.

AbFlex® Histone H3K4me3 antibody specificity is shown by Dot Blot.
Peptides corresponding to regions around major sites of histone H3 methylation (lysine 4, lysine 9, lysine 27) were spotted onto PVDF and probed with antibody at 1 µg/ml. The amount of peptide (in picomoles) spotted is indicated next to each row. Top panel - Lane 1: unmodified H3K4. Lane 2: H3K4me1. Lane 3: H3K4me2. Lane 4: H3K4me3. Lane 5: unmodified H3K9. Lane 6: H3K9me1. Lane 7: H3K9me2. Lane 8: H3K9me3. Lane 9: unmodified H3K79. Lane 10: H3K79me1. Lane 11: H3K79me2. Lane 12: H3K79me3. Bottom panel - Lane 1: unmodified H3K23. Lane 2: H3K23me1. Lane 3: H3K23me3. Lane 4: H3K23me3. Lane 5: unmodified H3K27. Lane 6: H3K27me1. Lane 7: H3K27me2. Lane 8: H3K27me3. Lane 9: unmodified H3K36. Lane 10: H3K36me1. Lane 11: H3K36me2. Lane 12: H3K36me3.

for AbFlex Histone H3K4me3 antibody (rAb)Background

AbFlex® antibodies are recombinant antibodies (rAbs) that have been generated using defined DNA sequences to produce highly specific, reproducible antibodies. Each AbFlex antibody contains a 6XHis tag, an avidin tag sequence for enzymatic biotin conjugation using the biotin ligase, BirA, and a sortase recognition motif (LPXTG) to attach a variety of labels directly to the antibody including fluorophores, enzymatic substrates (HRP, AP), peptides, drugs as well as solid supports. AbFlex H3K4me3 antibody was expressed as full-length IgG with mouse immunoglobulin heavy and light chains (IgG2a isotype) in mammalian 293 cells.

Histone H3 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). Histone H1 is a linker histone, present at the interface between the nucleosome core and DNA entry/exit points. Histone H1 is responsible for establishing higher-order chromatin structure. 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 methylation can be associated with transcriptional activation or repression, depending on the methylated residue. Lysine 4 of histone H3 can be mono-, di- or trimethylated by different histone methyltransferases (HMTs) such as SET1 or ASH1. Methylation of Lys4 is often associated with transcriptional activation. The demethylase LSD1 is able to demethylate histone H3 Lys4.

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 AbFlex® Histone H3K4me3 antibody (rAb).

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 AbFlex® Histone H3K4me3 antibody (rAb).

for AbFlex Histone H3K4me3 antibody (rAb)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 AbFlex Histone H3K4me3 antibody (rAb)Guarantee

This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.

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

AbFlex® Histone H3K4me3 antibody (rAb)
 

Data Thumbnails

AbFlex<sup>®</sup> Histone H3K4me3 antibody (rAb) tested by ChIP-Seq.

ChIP-Seq of AbFlex® Histone H3K4me3 antibody (rAb).
(Click image to enlarge and see details).

AbFlex<sup>®</sup> Histone H3K4me3 antibody (rAb) tested by Western blot.

Western blot of AbFlex® Histone H3K4me3 antibody (rAb).
(Click image to enlarge and see details.)

AbFlex<sup>®</sup> Histone H3K4me3 antibody (rAb) tested by dot blot analysis.

Dot blot of AbFlex® Histone H3K4me3 antibody (rAb).
(Click image to enlarge and see details.)