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Antibodies

Histone H3K4me3 antibody (pAb)

Catalog No: 39159 Format: 100 µl $420 Buy Now
Catalog No: 39160 Format: 10 µl $95 Buy Now

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Antibody Type:Polyclonal
Isotype:Serum
Purification:None
Host:Rabbit
Molecular Weight:17 kDa
Reactivity:Budding Yeast, Human, Mouse, Other (Wide Range)

Applications

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

for Histone H3 trimethyl Lys4 antibody (pAb)Application Notes


Validated Applications:
ChIP: 3 - 5 µl per ChIP
ChIP-Seq: 3 µl each
ICC/IF: 1:500 - 1:1,000 dilution
WB: 1:1,000 - 1:5,000 dilution

modENCODE validation. This antibody was validated for ChIP-Seq in this study (see reference).
NGS-QC® certification. This antibody has been processed by the NGS-QC® generator. For additional details, click here..

Published Applications:
ChIP, ChIP-chip, & ChIP-Seq
WB
IF
FC
See references for more information. Individual optimization may be required.

for Histone H3 trimethyl Lys4 antibody (pAb)Immunogen

This Histone H3 trimethyl Lys4 antibody was raised against a peptide including trimethyl-lysine 4 of histone H3.

for Histone H3 trimethyl Lys4 antibody (pAb)Buffer

Rabbit serum containing 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic. For your convenience, an IgG version (Catalog No. 39915) of this antibody that was purified by Protein A Chromatography is also available.

for Histone H3 trimethyl Lys4 antibody (pAb)References

 
Histone H3K4me3 antibody (pAb) tested by ChIP-Seq.

Histone H3 trimethyl Lys4 antibody tested by ChIP-Seq.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with chromatin from 2 million HL-60 cells and 3 µl of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 19 million sequence tags were mapped to identify H3K4me3 binding sites. The image shows H3K4me3 bound to most promoters of genes in the ZNF gene cluster on chromosome 19. (Gene annotations run from right to left, therefore TSSs are on the right side of each gene annotation.)

Histone H3K4me3 antibody (pAb) tested by ChIP.

Histone H3 trimethyl Lys4 antibody (pAb) tested by ChIP.
Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 2 million HL-60 cells and 3 µl of Histone H3 trimethyl Lys4 antibody. ChIP DNA was used in qPCR with the negative control primer pairs or gene-specific primer pairs as indicated. Data are presented as Binding Events Detected per 1000 Cells using Active Motif's Epigenetic Services normalization scheme which accounts for primer efficiency and the amount of chromatin used in the ChIP reaction.

Histone H3K4me3 antibody (pAb) tested by immunofluorescence.

Histone H3 trimethyl Lys4 antibody tested by immunofluorescence.
HeLa cells stained at 1:1,000 with Histone H3 trimethyl Lys4 antibody. Top left: Histone H3 trimethyl Lys4 antibody.Top right: Stained with DAPI. Bottom left: Same cells stained with alpha Tubulin mAb (Clone 5-B-1-2). Bottom right: Merge of all 3 images.

Histone H3K4me3 antibody (pAb) tested by Western blot.

Histone H3 trimethyl Lys4 antibody tested by Western blot.
HeLa nuclear extract (20 µg) was probed with Histone H3 trimethyl Lys4 antibody (1:2,000 dilution).

Histone H3K4me3 antibody (pAb) tested by dot blot analysis.

Histone H3 trimethyl Lys4 antibody tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 trimethyl Lys4 antibody for trimethyl-lysine 4 of histone H3. Peptides corresponding to regions around major sites of histone H3 methylation (lysine 4, lysine 9, lysine 27) were spotted onto PVDF and probed with the antibody at a dilution of 1:5,000. 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 Histone H3 trimethyl Lys4 antibody (pAb)Background

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.

for Histone H3 trimethyl Lys4 antibody (pAb)Positive Control

Active Motif's HeLa acid extract (Catalog No. 36200) can be used as a positive control for Histone H3 trimethyl Lys4 antibody.

for Histone H3 trimethyl Lys4 antibody (pAb)qPCR Primers

Active Motif's Human Positive Control Primer Set ACTB-2 (Cat. No. 71005) and Human Negative Control Primer Set 1 (Cat. No. 71001) have been validated for performing qPCR and endpoint PCR when this antibody has been used for ChIP on human samples. For ChIP experiments with this antibody on mouse samples, the Mouse Positive Control Primer Set Actb-2 (Cat. No. 71017) and Mouse Negative Control Primer Set 1 (Cat. No. 71011) have been validated for both qPCR and endpoint PCR.

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 H3K4me3 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 H3K4me3 antibody (pAb).

for Histone H3 trimethyl Lys4 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.

for Histone H3 trimethyl Lys4 antibody (pAb)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 H3K4me3 antibody (pAb)
 

Data Thumbnails

Histone H3K4me3 antibody (pAb) tested by ChIP-Seq.

ChIP-Seq of Histone H3K4me3 pAb.
(Click image to enlarge and see details.)

Histone H3K4me3 antibody (pAb) tested by ChIP.

ChIP of Histone H3K4me3 pAb.
(Click image to enlarge and see details.)

Histone H3K4me3 antibody (pAb) tested by immunofluorescence.

Immunofluorescence stain of Histone H3K4me3 pAb.
(Click image to enlarge and see details.)

Histone H3K4me3 antibody (pAb) tested by Western blot.

Western blot of Histone H3K4me3 pAb.
(Click image to enlarge and see details.)

Histone H3K4me3 antibody (pAb) tested by dot blot analysis.

Dot blot of Histone H3K4me3 pAb.
(Click image to enlarge and see details.)