推荐产品
生物源
rabbit
品質等級
抗體表格
culture supernatant
抗體產品種類
primary antibodies
無性繁殖
AW30, monoclonal
物種活性
human, chicken
製造商／商標名
Upstate®
技術
ChIP: suitable (ChIP-seq)
dot blot: suitable
inhibition assay: suitable (peptide)
western blot: suitable
同型
IgG
NCBI登錄號
UniProt登錄號
運輸包裝
wet ice
目標翻譯後修改
dimethylation (Lys4)
基因資訊
human ... H3C1(8350)
一般說明
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.
特異性
Histone H3 dimethylated on lysine 4
The immunizing sequence is conserved from Tetrahymena to human, so broad species cross-reactivity is expected.
免疫原
KLH-conjugated, synthetic peptide containing the sequence …Tme2KQT… in which me2K corresponds to dimethyl-lysine at residue 4 of human histone H3.
應用
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-S), 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-S), 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
品質
routinely evaluated by immunoblot on acid-extracted proteins from HeLa cells; the antibody did not detect unmethylated recombinant Histone H3 (Catalog #14-494)
聯結
Replaces: 05-790
外觀
Cultured supernantant in 0.05% sodium azide
儲存和穩定性
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.
分析報告
Control
HeLa acid extracts
HeLa acid extracts
法律資訊
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
免責聲明
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.
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儲存類別代碼
12 - Non Combustible Liquids
水污染物質分類（WGK）
WGK 1
閃點（°F）
Not applicable
閃點（°C）
Not applicable
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