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Histone H3 trimethyl Lys4 antibody (pAb)
| Catalog No: 39159 | Format: 200 µl | $340 | Buy Now |
| Catalog No: 39160 | Format: 10 µl | $70 | Buy Now |
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Applications Table
| Western Blot | Chromatin IP | ChIP-Seq / ChIP-chip |
IF |
|---|---|---|---|
| 1:1,000 - 1:5,000 | 3 - 5 µl per ChIP | 3 µl per ChIP-Seq 3 µl per ChIP-chip |
1:500 - 1:1,000 |
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.
The methylation of histones can occur on two different residues: arginine or lysine. 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)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)Application Notes
This Histone H3 trimethyl Lys4 antibody (H3K4me3) has been validated for use in chromatin IP (ChIP, 3 - 5 µl), ChIP-Seq & ChIP-chip (3 µl each), immunofluorescence (1:500 - 1:1,000) and Western blotting (1:1,000 - 1:5,000 dilution). Specificity for trimethyl lysine 4 was confirmed by dot blot with non-modified histone H3 peptide or peptides mono-, di- or trimethylated at lysine 4; peptides mono-, di- or trimethylated at lysine 9; peptides mono-, di- or trimethylated at lysine 27. ChIP using a yeast strain with a point mutation at lysine 4 was negative. Also, Western blot against 200 ng of recombinant histone H3 was negative. Because this modification is conserved from yeast to human, a wide range of reactivity is expected. This antibody is specific for histone H3 trimethylated at lysine 4 (see dot blot data below).
The modENCODE and NIH Roadmap Epigenomics Mapping Consortiums have implemented rigorous standardization criteria for all assays and reagents to be used. As part of this initiative, antibody specificity testing and the ability of the antibodies to work in ChIP-Seq were assessed in a large-scale study. This Histone H3 trimethyl Lys4 antibody was validated for ChIP-Seq in the study (see References).
for Histone H3 trimethyl Lys4 antibody (pAb)Buffer
Rabbit serum containing 30% glycerol and 0.035% sodium azide.
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.
Chromatin IP Assays
Active Motif has a variety of Chromatin Immunoprecipitation Kits for every need.
Please visit our Chromatin IP Page to learn about ChIP products designed for use with Histone H3 trimethyl Lys4 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 H3 trimethyl Lys4 antibody (pAb).
for Histone H3 trimethyl Lys4 antibody (pAb)References
This antibody has been used as described below:
Histone H3 trimethyl Lys4 antibody tested by Western blot.
HeLa acid extract (10 µg) was probed with Histone H3 trimethyl Lys4 antibody (1:1,000 dilution).
Histone H3 trimethyl Lys4 antibody tested by ChIP.
Top Panel: ChIP with wild-type yeast chromatin (lanes 1 and 3) or a strain containing a point mutation at lysine 4 (lane 2) using 3 µl Histone H3 trimethyl Lys4 antibody (Catalog No. 39159) (lanes 1 and 2) or 3 µl Histone H3 acetyl Lys27 rabbit antibody (Catalog No. 39133) (lane 3). Primers specific to a region of the SCC1 gene promoter were used to analyze the products by PCR.
Bottom Panel: Chromatin immunoprecipitation performed using the ChIP-IT® Express Kit (Catalog No. 53008) and 25 µl of Ready-to-ChIP HeLa Chromatin (Catalog No. 53015) using 5 µl of Histone H3 trimethyl Lys4 antibody or the equivalent amount of rabbit IgG as a negative control. RT-qPCR was performed on ChIP DNA using a primer pair specific for the indicated gene. Data are presented as Fold Enrichment of the ChIP antibody signal versus the negative control IgG using the ddCT method.
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.
Lane 1: Unmodified Lys4 peptide. Lane 2: Monomethyl Lys4 peptide. Lane 3: Dimethyl Lys4 peptide. Lane 4: Trimethyl Lys4 peptide. Lane 5: Monomethyl Lys9 peptide. Lane 6: Unmodified Lys9 peptide. Lane 7: Dimethyl Lys9 peptide. Lane 8: Trimethyl Lys9 peptide. Lane 9: Unmodified Lys27 peptide. Lane 10: Monomethyl Lys27 peptide. Lane 11: Dimethyl Lys27 peptide. Lane 12: Trimethyl Lys27 peptide.
Histone H3 trimethyl Lys4 antibody tested by immunofluorescence.
HeLa cells stained at 1:500 with Histone H3 trimethyl Lys4 antibody. Top panel: Histone H3 trimethyl Lys4 antibody. Middle panel: DAPI. Bottom panel: merge.
Histone H3 trimethyl Lys4 antibody tested by ChIP-Seq.
ChIP was performed using chromatin from 500,000 primary mouse T cells. ChIP DNA was sequenced on the Illumina GA II and 25 million sequence tags were mapped to identify H3K4me3 binding sites. The image shows H3K4me3 bound to the promoters of all genes in the Zfp gene cluster on chromosome 13. (Gene annotations run from right to left, therefore TSSs are on the right side of each gene annotation.)
for Histone H3 trimethyl Lys4 antibody (pAb)Storage
Antibodies in solution can be stored at -20°C for 2 years. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
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
- FACS = Flow Cytometry;
- ICC = Immunocytochemistry;
- IF = Immunofluorescence;
- IHC = Immunohistochemistry;
- IP = Immunoprecipitation;
- MeDIP = Methyl-DNA Immunoprecipitation;
- WB = Western Blotting
