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ABE250 Anti-dimethyl Histone H3 (Lys4) Antibody

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ABE250
100 µL  
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Overview

Replacement Information

Key Spec Table

Species ReactivityKey ApplicationsHostFormatAntibody Type
H, M, R, ChWB, ICC, DB, ChIP, ChIP-seqRbAffinity PurifiedPolyclonal Antibody
Description
Catalogue NumberABE250
Replaces04-791
DescriptionAnti-dimethyl Histone H3 (Lys4) Antibody
Alternate Names
  • H3 histone family, member T
  • histone 3, H3
  • histone cluster 3, H3, H3K4me2
Background InformationHistone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 is involved with the structure of the nucleosomes of the 'beads on a string' structure. The N-terminal tail of histone H3 protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine. Dimethylation of histone H3 at Lys4 correlates with transcriptional activity of many genes.
References
Product Information
FormatAffinity Purified
Control
  • HeLa acid extract
PresentationPurified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Quality LevelMQ100
Applications
ApplicationAnti-dimethyl Histone H3 (Lys4) Antibody is a Rabbit Polyclonal Antibody for detection of Histone H3 dimethylated at lysine 4. This purified polyclonal antibody, also known as H3K4me2, has dot blot (DB) proven specificity & has been validated in WB, ICC, ChIP.
Key Applications
  • Western Blotting
  • Immunocytochemistry
  • Dot Blot
  • Chromatin Immunoprecipitation (ChIP)
  • ChIP-seq
Application NotesImmunocytochemistry Analysis:
1:500 dilution from a representative lot detected Histone H3 in NIH/3T3 and A431 cells.

Dot Blot Analysis:
1:1,000 dilution from a representative lot specifically detected Histone H3 in a Lys4 dimethhyl modified histone H3 peptide while not detecting potential cross-reacting peptides corresponding to other modified and non-modified histones.

Chromatin Immunoprecipitation Analysis:
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 4 µg of either Normal IgG (Part No. 12-370), or 4 µL of Anti-Dimethyl Histone H3 (Lys4) (Part No. ABE250) and the Magna ChIP™ A/G Kit (Cat. # 17-10085). Successful immunoprecipitation of Dimethyl-Histone H3 (Lys4) associated DNA fragments was verified by qPCR using primers specific for the human GAPDH coding region.
Please refer to the EZ-Magna ChIP™ A/G (Cat. # 17-10085) protocol for experimental details.

ChIP-Sequencing
Chromatin immunoprecipitation was performed using the Magna ChIP™ HiSens kit (cat# 17-10460), 5 µg ofa representative lot of anti-dimethyl-Histone H3 (Lys4) antibody(ABE250), 20 µL Protein A/G beads ,and 5e6 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 twelve 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.
Biological Information
ImmunogenKLH-conjugated linear peptide corresponding Histone H3 methylated at Lys4.
EpitopeDimethylated Lys4
HostRabbit
SpecificityThis antibody recognizes Histone H3 dimethylated at Lys4.
Species Reactivity
  • Human
  • Mouse
  • Rat
  • Chicken
Species Reactivity NoteDemonstrated to react with human, mouse, rat, and chicken. Broad species cross-reactivity expected based on sequence similarity.
Antibody TypePolyclonal Antibody
Entrez Gene Number
Entrez Gene SummaryHistones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is located separately from the other H3 genes that are in the histone gene cluster on chromosome 6p22-p21.3. [provided by RefSeq].
Gene Symbol
  • HIST1H3A
  • H3FA
  • HIST1H3B
  • H3FL
  • HIST1H3C
  • H3FC
  • HIST1H3D
  • H3FB
  • HIST1H3E
  • H3FD
  • HIST1H3F
  • H3FI
  • HIST1H3G
  • H3FH
  • HIST1H3H
  • H3FK
  • HIST1H3I
  • H3FF
  • HIST1H3J
  • H3FJ
  • HIST3H3
  • H3FT
  • H3T
  • MGC126886
Modifications
  • Methylation
Purification MethodAffinity Purfied
UniProt Number
UniProt SummaryFUNCTION: Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.

SIZE: 136 amino acids; 15508 Da

SUBUNIT: The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA.

SUBCELLULAR LOCATION: Nucleus.

TISSUE SPECIFICITY: Expressed in testicular cells.

DEVELOPMENTAL STAGE: Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.

PTM: Acetylation is generally linked to gene activation. Acetylation on Lys-10 impairs methylation at Arg-9. Acetylation on Lys-19 and Lys-24 favors methylation at Arg-18 (By similarity). & Citrullination at Arg-9 and/or Arg-18 by PADI4 impairs methylation and represses transcription (By similarity). & Asymmetric dimethylation at Arg-18 by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 by PRMT5 is linked to gene repression (By similarity). & Methylation at Lys-5, Lys-37 and Lys-80 are linked to gene activation. Methylation at Lys-5 facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 and Lys-28 are linked to gene repression. Methylation at Lys-10 is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 and acetylation of H3 and H4. Methylation at Lys-5 and Lys-80 require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 and Lys-28 are enriched in inactive X chromosome chromatin (By similarity). & Phosphorylated at Thr-4 by GSG2/haspin during prophase and dephosphorylated during anaphase. At centromeres, specifically phosphorylated at Thr-12 from prophase to early anaphase. Phosphorylated at Ser-11 during the whole mitosis. Phosphorylation at Ser-11, which is linked to gene activation, prevents methylation at Lys-10 but facilitates acetylation of H3 and H4. Phosphorylated at Ser-29 by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation (By similarity). & Phosphorylation at 'Ser-11' is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at 'Ser-11' is important during interphase because it enables the transcription of genes following external stimulation, like stress or growth factors. Phosphorylation at 'Ser-11' is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylation at 'Ser-11' by AURKB/Aurora-B mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. & Ubiquitinated (By similarity).

SIMILARITY: Belongs to the histone H3 family.
Molecular Weight~17 kDa observed
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality AssuranceEvaluated by Western Blot using HeLa acid extract and recombinant histone proteins.

Western Blot Analysis: 1:1,000 dilution of this antibody detected Histone H3 on 10 µg of HeLa acid extract.
Usage Statement
  • 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.
Storage and Shipping Information
Storage ConditionsStable for 1 year at 2-8°C from date of receipt.
Packaging Information
Material Size100 µL
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
ABE250 04053252554544

Documentation

Anti-dimethyl Histone H3 (Lys4) Antibody SDS

Title

Safety Data Sheet (SDS) 

Anti-dimethyl Histone H3 (Lys4) Antibody Certificates of Analysis

TitleLot Number
Anti-dimethyl Histone H3 (Lys4) 3076545
Anti-dimethyl Histone H3 (Lys4) - 2377656 2377656
Anti-dimethyl Histone H3 (Lys4) - 2445049 2445049
Anti-dimethyl Histone H3 (Lys4) - 2074779 2074779
Anti-dimethyl Histone H3 (Lys4) - 2172765 2172765
Anti-dimethyl Histone H3 (Lys4) - 2204965 2204965
Anti-dimethyl Histone H3 (Lys4) - 2283570 2283570
Anti-dimethyl Histone H3 (Lys4) - 3239891 3239891
Anti-dimethyl Histone H3 (Lys4) - 4035058 4035058
Anti-dimethyl Histone H3 (Lys4) - NG1905723 NG1905723

References

Reference overviewPub Med ID
Cytomixis doesn't induce obvious changes in chromatin modifications and programmed cell death in tobacco male meiocytes.
Mursalimov, S; Permyakova, N; Deineko, E; Houben, A; Demidov, D
Frontiers in plant science  6  846  2015

Show Abstract
26528310 26528310
Modulation of chromatin remodelling induced by the freshwater cyanotoxin cylindrospermopsin in human intestinal caco-2 cells.
Huguet, A; Hatton, A; Villot, R; Quenault, H; Blanchard, Y; Fessard, V
PloS one  9  e99121  2014

Show Abstract
24921660 24921660

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Life Science Research > Antibodies and Assays > Primary Antibodies