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05-1338-S Anti-Dimethyl Histone H3 (Lys4) Antibody, clone CMA303, Trial Size

Anti-Dimethyl Histone H3 (Lys4) Antibody, clone CMA303 is a mouse monoclonal antibody for detection of Dimethyl Histone H3 (Lys4) also known as H3K4me2, Histone H3 (di methyl K4). Demonstrated performance in ChIP, DB, ELISA, ICC, IP, WB, Mplex.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Research Category: Epigenetics & Nuclear Function Research Sub-Category: Histones Gene Symbol: H3.4, H3/g , H3/t , H3FT , H3T, H3t , MGC126886, MGC126888 , OTTHUMP00000037945 UniProt Number: Q16695
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05-1338-S
10 µg  
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Key Specifications Table

Species ReactivityKey ApplicationsHostFormatAntibody Type
H, VrtWB, ChIP, ICC, IP, Mplex, DB, ELISAMPurifiedMonoclonal Antibody
Description
Catalogue Number05-1338-S
DescriptionAnti-Dimethyl Histone H3 (Lys4) Antibody, clone CMA303, Trial Size
Alternate Names
  • H3K4me2
  • Histone H3 (dimethyl K4)
  • H3 histone family, member T
  • Histone 3, H3
  • Histone cluster 3, H3
Background InformationHistones 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. Histones are modified post-translationally; and these modifications regulate DNA transcription, repair, recombination, and replication. The most commonly studied modifications are acetylation, phosphorylation, methylation, and ubiquitination. The modifications occur predominantly on the N-terminal and C-terminal tails that extend beyond the nucleosome core particle. Dimethyl-lysine 4 histone H3 (H3K4me2) is a transcription-activating chromatin mark, and dimethyl histone H3 Lys4 (H3K4me2) is depleted from regions with DNA methylation. Multipotential hematopoietic cells have a subset of genes that are differentially methylated (H3K4me2+/me3-). These genes are transcriptionally silent and highly enriched in lineage-specific hematopoietic genes, suggesting a role for H3K4 methylation in differentiation.
References
Product Information
FormatPurified
PresentationPurified mouse monoclonal IgG1κ in PBS with 0.05% sodium azide.
Quality LevelMQ100
Applications
ApplicationAnti-Dimethyl Histone H3 (Lys4) Antibody, clone CMA303 is a mouse monoclonal antibody for detection of Dimethyl Histone H3 (Lys4) also known as H3K4me2, Histone H3 (di methyl K4). Demonstrated performance in ChIP, DB, ELISA, ICC, IP, WB, Mplex.
Key Applications
  • Western Blotting
  • Chromatin Immunoprecipitation (ChIP)
  • Immunocytochemistry
  • Immunoprecipitation
  • Multiplexing
  • Dot Blot
  • ELISA
Application NotesChromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either a normal mouse IgG, or Anti-dimethyl-Histone H3 (Lys4) antibody and the Magna ChIP G (Cat. # 17-611) Kit. Successful immunoprecipitation of dimethyl-histone H3 (Lys4) associated DNA fragments was verified by qPCR using ChIP Primers GAPDH Coding.
Please refer to the EZ-Magna G ChIP™ (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

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 05-1338-S, Anti-dimethyl H3 (Lys4), clone CMA303 at 2.0 µg/mL (1:500 dilution). Proteins were visualized using a Donkey anti-mouse IgG conjugated to HRP and a chemiluminescence detection system.

ChIP-seq Analysis:
Chromatin immunoprecipitation was performed using the Magna ChIP™ HiSens kit (cat# 17-10460), 2 µg of Anti-dimethyl-Histone H3 (Lys4) antibody (cat# 05-1338-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.

Immunocytochemistry:
This antibody has been shown by an outside laboratory to be suitable for immunocytochemistry.

Immunoprecipitation:
This antibody has been shown by an outside laboratory to be suitable for immunoprecipitation.

ELISA:
This antibody has been shown by an outside laboratory to be suitable for ELISA

Multiplexing:
This antibody specifically recognizes histone H3 dimethylated on Lys4 by Luminex® assay.
Biological Information
ImmunogenSynthetic peptide corresponding to amino acids 1-12 of human Histone H3, dimethylated on Lys4, conjugated to KLH.
EpitopeDimethylated Lys4
CloneCMA303
ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
HostMouse
SpecificityThis antibody specifically recognizes Histone H3 dimethylated at Lys4. The antibody binding specificity allows for phosphorylation of Thr3 and/or modifications of Lys91.
IsotypeIgG1κ
Species Reactivity
  • Human
  • Vertebrates
Species Reactivity NoteHuman. Broad species cross-reactivity is expected, based on sequence homology.
Antibody TypeMonoclonal 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.
Gene Symbol
  • H3.4
  • H3/g
  • H3/t
  • H3FT
  • H3T
  • H3t
  • MGC126886
  • MGC126888
  • OTTHUMP00000037945
Modifications
  • Methylation
Purification MethodProtein G Purified
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.

SUBUNIT STRUCTURE: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 Weightapprox. 17 kDa
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality AssuranceRoutinely evaluated by Western Blot on HeLa acid extract.

Western Blot Analysis: 2 μg/mL (1:500) dilution of this antibody detected dimethyl Histone H3 (Lys4) 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. For maximum recovery of product, centrifuge the vial prior to removing the cap. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Packaging Information
Material Size10 µg
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalog Number GTIN
05-1338-S 04053252929892

Documentation

Anti-Dimethyl Histone H3 (Lys4) Antibody, clone CMA303, Trial Size SDS

Title

Safety Data Sheet (SDS) 

Anti-Dimethyl Histone H3 (Lys4) Antibody, clone CMA303, Trial Size Certificates of Analysis

TitleLot Number
Anti-dimethyl Histone H3 (Lys4), -2623696 2623696
Anti-dimethyl Histone H3 (Lys4), -2759642 2759642
Anti-dimethyl Histone H3 (Lys4), -2770273 2770273
Anti-dimethyl Histone H3 (Lys4), -2780422 2780422
Anti-dimethyl Histone H3 (Lys4), clone CMA303, Trial Size - 3221499 3221499
Anti-dimethyl Histone H3 (Lys4), clone CMA303, Trial Size - 3693872 3693872
Anti-dimethyl Histone H3 (Lys4), clone CMA303, Trial Size - 4067244 4067244
Anti-dimethyl Histone H3 (Lys4), clone CMA303, Trial Size - 4160344 4160344
Anti-dimethyl Histone H3 (Lys4), clone CMA303, Trial Size - Q2290318 Q2290318
Anti-dimethyl Histone H3 (Lys4), clone CMA303, Trial Size Monoclonal Antibody 2910084

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Life Science Research > Antibodies and Assays > Primary Antibodies