Your Cart
Browse
Filter
Histone H3 monomethyl Lys4 antibody (pAb)
| Catalog No: 39297 | Format: 200 µl | $340 | Buy Now |
| Catalog No: 39298 | Format: 10 µl | $70 | Buy Now |
|
|
Applications Table
| Western Blot | Chromatin IP | ChIP-Seq / ChIP-chip |
IF |
|---|---|---|---|
| 1:5,000 - 1:20,000 | 10 µl per ChIP | 10 µl per ChIP-Seq 10 µl per ChIP-chip |
1:500 - 1:1,000 |
for Histone H3 monomethyl 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 monomethyl Lys4 antibody (pAb)Immunogen
This Histone H3 monomethyl Lys4 antibody was raised against a peptide including monomethyl-lysine 4 of human histone H3.
for Histone H3 monomethyl Lys4 antibody (pAb)Application Notes
This Histone H3 monomethyl Lys4 antibody (H3K4me1) has been validated for use in chromatin IP (10 µl per ChIP reaction), ChIP-Seq & ChIP-chip (10 µl each), Western blotting (1:5,000 - 1:20,000 dilution) and immunofluorescence (1:500 - 1:1,000 dilution).
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 monomethyl Lys4 antibody was validated for ChIP-Seq in the study (see References).
for Histone H3 monomethyl Lys4 antibody (pAb)Buffer
Rabbit serum containing 30% glycerol and 0.035% sodium azide.
for Histone H3 monomethyl Lys4 antibody (pAb)Positive Control
Active Motif's HeLa acid extract (Catalog No. 36200) can be used as a positive control for Histone H3 monomethyl 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 monomethyl 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 monomethyl Lys4 antibody (pAb).
for Histone H3 monomethyl Lys4 antibody (pAb)References
This antibody has been used as described below:
Histone H3 monomethyl Lys4 antibody tested by Western blot.
HeLa acid extract probed with Histone H3 monomethyl Lys4 antibody (1:5,000 dilution).
Histone H3 monomethyl Lys4 antibody tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 monomethyl Lys4 antibody for monomethyl-Lys4 histone H3. Methylated peptides corresponding to the immunogen and related peptides were spotted onto PVDF and probed with the antibody at 1:10,000. The amount of peptide (picomoles) spotted is indicated next to each row.
Lane 1: Unmodified lysine 4 peptide. Lane 2: Monomethyl lysine 4 peptide. Lane 3: Dimethyl lysine 4 peptide. Lane 4: Trimethyl lysine 4 peptide. Lane 5: Monomethyl lysine 4 peptide #2. Lane 6: Unmodified peptide corresponding to amino acids 6-22 of human histone H3. Lane 7: Monomethyl lysine 9 peptide. Lane 8: Dimethyl lysine 9 peptide. Lane 9: Trimethyl lysine 9 peptide. Lane 10: Dimethyl lysine 9 peptide #2. Lane 11: Unmodified lysine 27 peptide. Lane 12: Monomethyl lysine 27 peptide. Lane 13: Dimethyl lysine 27 peptide. Lane 14: Trimethyl lysine 27 peptide. Lane 15: Monomethyl lysine 27 peptide #2. Lane 16: Dimethyl lysine 14 peptide.
Histone H3 monomethyl Lys4 antibody (pAb) tested by immunofluorescence.
HeLa cells stained with Histone H3 monomethyl Lys4 antibody (1:1,000). Top: Histone H3 monomethyl Lys4 antibody. Middle: DAPI. Bottom: merge.
Histone H3 monomethyl Lys4 antibody specificity tested by peptide array analysis.
Peptide array analysis was used to confirm the specificity of this antibody for its intended modification. Histone H3 monomethyl Lys4 antibody was applied at a dilution of 1:5,000 to Active Motif's MODified™ Histone Peptide Array (Catalog No. 13001). The arrays were scanned with ArrayAnalysis Software 7 and the results plotted. Specificity data is shown for the most reactive peptides and those related to the immunogen. Recognition of the H3 monomethyl Lys4 peptide is blocked by Thr3 phosphorylation, Arg2 methylation and a citrulline at position 2.
Array Data File
Histone H3 trimethyl Lys4 antibody tested by ChIP-Seq.
ChIP was performed using human prostate cancer cells ( 3 million). ChIP DNA was sequenced on the Illumina GA II and 22 million sequence tags were mapped to identify H3K4me1 binding sites. The image shows H3K4me1 binding across a 600 Kb region on chromosome 13.
for Histone H3 monomethyl 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 monomethyl 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
