04-790
Anti-dimethyl-Histone H3 (Lys4) Antibody, clone AW30, rabbit monoclonal
culture supernatant, clone AW30, Upstate®
Synonym(s):
H3K4me2, Histone H3 (di methyl K4)
Sign Into View Organizational & Contract Pricing
All Photos(5)
About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41
Recommended Products
biological source
rabbit
Quality Level
antibody form
culture supernatant
antibody product type
primary antibodies
clone
AW30, monoclonal
species reactivity
chicken, human
manufacturer/tradename
Upstate®
technique(s)
ChIP: suitable (ChIP-seq)
dot blot: suitable
inhibition assay: suitable (peptide)
western blot: suitable
isotype
IgG
NCBI accession no.
UniProt accession no.
shipped in
dry ice
target post-translational modification
dimethylation (Lys4)
Gene Information
human ... H3C1(8350)
General description
Histones are highly conserved proteins that serve as the structural scaffold for the organization of nuclear DNA into chromatin. The four core histones, H2A, H2B, H3, and H4, assemble into an octamer (2 molecules of each). Subsequently, 146 base pairs of DNA are wrapped around the octamer, forming a nucleosome, the basic subunit of chromatin. Histones are modified post-translationally by the actions of enzymes in both the nucleus and cytoplasm. These modifications regulate DNA transcription, repair, recombination, and replication. The most commonly studied modifications are acetylation, phosphorylation, methylation, and ubiquitination. These modifications occur predominantly on the N-terminal and C-terminal tails that extend beyond the nucleosome core particle. Di- and trimethylation of histone H3 at Lys4 correlates with transcriptional activity of many genes. Dimethylation of Histone H3 at Lys4 occurs at both active and inactive euchromatic regions but not in silent heterochromatic sites, whereas trimethylation at Lys4 is present exclusively at active genes.
Specificity
Histone H3 dimethylated on lysine 4
The immunizing sequence is conserved from Tetrahymena to human, so broad species cross-reactivity is expected.
Immunogen
KLH-conjugated, synthetic peptide containing the sequence …Tme2KQT… in which me2K corresponds to dimethyl-lysine at residue 4 of human histone H3.
Application
Western Blot Analysis:
A 1:1000-1:2000 dilution of this lot detected dimethyl histone H3 (Lys4) in HeLa acid extracts.
Western Blot Analysis:
A 1:2,000 to 1:10,000 dilution of a previous lot detected methylated histone H3 in acid extracted proteins from HeLa cells. The antibody did not detect unmethylated recombinant Histone H3.
ChIP-seq Analysis:
Representative lot data. Chromatin immunoprecipitation was performed using the Magna ChIP HiSens kit (cat# 17-10460), Anti-dimethyl-Histone H3 (Lys4) antibody (2 µl cat# 04-790), 20 µL Protein A/G beads, and 1e6 crosslinked HeLa cell chromatin followed by DNA purification using magnetic beads. Libraries were prepared from Input and ChIP DNA samples using standard protocols with Illumina barcoded adapters, and analyzed on Illumina HiSeq instrument. An excess of sixteen million reads from FastQ files were mapped using Bowtie (http://bowtie-bio.sourceforge.net/manual.shtml) following TagDust (http://genome.gsc.riken.jp/osc/english/dataresource/) tag removal. Peaks were identified using MACS (http://luelab.dfci.harvard.edu/MACS/), with peaks and reads visualized as a custom track in UCSC Genome Browser (http://genome.ucsc.edu) from BigWig and BED files. The highest 25% of peaks identified in the 04-790 and 05-1338 datasets showed 92 and 90% overlap with peaks identified in the ENCODE H3K4me2 BROAD Histone track for HeLa S3.
Dot Blot Analysis:
Absurance Histone H3 Antibody Specificity Array (Cat. No. 16-667) and Absurance Histone H2A, H2B, H4 Antibody Specificity Array (Cat. No. 16-665), which contain histone peptides with various modifications were probed with Cat. No 04-790, Anti-dimethyl Histone H3 (Lys4), clone AW30 at 1:1000 dilution. Proteins were visualized using a Donkey anti-Rabbit IgG conjugated to HRP and a chemiluminescence detection system.
Peptide Inhibition Analysis:
2 μM of a histone H3 peptide containing dimethyl-lysine 4 abolished detection of histone H3 by a previous lot of antibody in immunoblot analysis of HeLa acid extracts. No signal reduction was observed with histone H3 peptides containing either monomethyl or trimethyl-lysine 4 modifications
A 1:1000-1:2000 dilution of this lot detected dimethyl histone H3 (Lys4) in HeLa acid extracts.
Western Blot Analysis:
A 1:2,000 to 1:10,000 dilution of a previous lot detected methylated histone H3 in acid extracted proteins from HeLa cells. The antibody did not detect unmethylated recombinant Histone H3.
ChIP-seq Analysis:
Representative lot data. Chromatin immunoprecipitation was performed using the Magna ChIP HiSens kit (cat# 17-10460), Anti-dimethyl-Histone H3 (Lys4) antibody (2 µl cat# 04-790), 20 µL Protein A/G beads, and 1e6 crosslinked HeLa cell chromatin followed by DNA purification using magnetic beads. Libraries were prepared from Input and ChIP DNA samples using standard protocols with Illumina barcoded adapters, and analyzed on Illumina HiSeq instrument. An excess of sixteen million reads from FastQ files were mapped using Bowtie (http://bowtie-bio.sourceforge.net/manual.shtml) following TagDust (http://genome.gsc.riken.jp/osc/english/dataresource/) tag removal. Peaks were identified using MACS (http://luelab.dfci.harvard.edu/MACS/), with peaks and reads visualized as a custom track in UCSC Genome Browser (http://genome.ucsc.edu) from BigWig and BED files. The highest 25% of peaks identified in the 04-790 and 05-1338 datasets showed 92 and 90% overlap with peaks identified in the ENCODE H3K4me2 BROAD Histone track for HeLa S3.
Dot Blot Analysis:
Absurance Histone H3 Antibody Specificity Array (Cat. No. 16-667) and Absurance Histone H2A, H2B, H4 Antibody Specificity Array (Cat. No. 16-665), which contain histone peptides with various modifications were probed with Cat. No 04-790, Anti-dimethyl Histone H3 (Lys4), clone AW30 at 1:1000 dilution. Proteins were visualized using a Donkey anti-Rabbit IgG conjugated to HRP and a chemiluminescence detection system.
Peptide Inhibition Analysis:
2 μM of a histone H3 peptide containing dimethyl-lysine 4 abolished detection of histone H3 by a previous lot of antibody in immunoblot analysis of HeLa acid extracts. No signal reduction was observed with histone H3 peptides containing either monomethyl or trimethyl-lysine 4 modifications
Anti-dimethyl-Histone H3 (Lys4) Antibody, clone AW30 is a rabbit monoclonal antibody for detection of dimethyl-Histone H3 (Lys4) also known as H3K4me2, Histone H3 (di methyl K4) & has been validated in WB, ChIP, ChIP-seq, DB, PIA, Mplex.
Research Category
Epigenetics & Nuclear Function
Epigenetics & Nuclear Function
Research Sub Category
Histones
Histones
Quality
routinely evaluated by immunoblot on acid-extracted proteins from HeLa cells; the antibody did not detect unmethylated recombinant Histone H3 (Catalog #14-494)
Target description
~17kDa
Linkage
Replaces: 05-790
Physical form
Cultured supernantant in 0.05% sodium azide
Storage and Stability
Stable for 1 year at -20°C from date of receipt.
For maximum recovery of product, centrifuge the vial prior to removing the cap.
For maximum recovery of product, centrifuge the vial prior to removing the cap.
Analysis Note
Control
HeLa acid extracts
HeLa acid extracts
Legal Information
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Not finding the right product?
Try our Product Selector Tool.
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 1
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
Already Own This Product?
Find documentation for the products that you have recently purchased in the Document Library.
Barbara Meller et al.
Frontiers in plant science, 9, 1228-1228 (2018-09-21)
In this paper we analyzed β-aminobutyric acid (BABA)-primed epigenetic adjustment of potato cv. "Sarpo Mira" to Phytophthora infestans. The first stress-free generation of the potato genotype obtained from BABA-primed parent plants via tubers and seeds showed pronounced resistance to the
A role for Set1/MLL-related components in epigenetic regulation of the Caenorhabditis elegans germ line.
Li, T; Kelly, WG
PLoS Genetics null
Structural basis of a histone H3 lysine 4 demethylase required for stem elongation in rice.
Chen, Q; Chen, X; Wang, Q; Zhang, F; Lou, Z; Zhang, Q; Zhou, DX
PLoS Genetics null
X Zhao et al.
Oncogene, 35(27), 3565-3576 (2015-11-10)
Previous studies have indicated the important roles of MYCN in tumorigenesis and progression of neuroblastoma (NB), the most common extracranial solid tumor derived from neural crest in childhood. However, the regulatory mechanisms of MYCN expression in NB still remain largely
Rachel Bayley et al.
Molecular cell, 82(10), 1924-1939 (2022-04-20)
The 53BP1-RIF1-shieldin pathway maintains genome stability by suppressing nucleolytic degradation of DNA ends at double-strand breaks (DSBs). Although RIF1 interacts with damaged chromatin via phospho-53BP1 and facilitates recruitment of the shieldin complex to DSBs, it is unclear whether other regulatory
Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.
Contact Technical Service