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17-10250
Sigma-AldrichChIPAb+™ Dimethyl-Histone H4 (Arg3) Symmetric - ChIP Validated Antibody and Primer Set
This ChIPAb+ Dimethyl-Histone H4 (Arg3) Symmetric -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
More>>This ChIPAb+ Dimethyl-Histone H4 (Arg3) Symmetric -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. Less<<
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ChIPAb+™ Dimethyl-Histone H4 (Arg3) Symmetric - ChIP Validated Antibody and Primer Set
Overview
All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction. The ChIPAb+ Dimethyl-Histone H4 (Arg3) Symmetric set includes the Dimethyl-Histone H4 (Arg3) Symmetric antibody, a Normal Rabbit IgG, and control primers which amplify a 110 bp region of ChIP Primers, human β-globin. The Dimethyl-Histone H4 (Arg3) Symmetric and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Dimethyl-Histone H4 (Arg3) Symmetric-associated chromatin.
Alternate Names
Histone H4
Background Information
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). Histones are modified post-translationally by the actions of enzymes in both the nucleus and cytoplasm. These modifications, which include acetylation, phosphorylation, methylation, and ubiquitin-ation, occur predominantly on the N-terminal and C-terminal tails that extend beyond the nucleosome core particle. Arginine modifications of histones proteins are mediated by two classes of protein arginine methyltransferases (PRMTs), class I and II. Class I PRMTs place two methyl groups on the one nitrogen atom of the arginine guanidino group, resulting in an asymmetric modification. In contrast class II PRMTs place two methyl groups on two nitrogen atoms of the guanidino group producing a symmetric dimethylation. Although there are exceptions, asymmetric modifications are typically associated with transcription activation while symmetric modifications are most commonly associated with transcriptional repression.
References
Product Information
Format
Affinity Purified
Control
Includes normal rabbit IgG and primers specific for human β-globin.
Presentation
Anti-Dimethyl-Histone H4 (Arg3) Symmetric (rabbit polyclonal). One vial containing 17.5 µg of purified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide before the addition of glycerol to 30%. Store at -20°C. Concentration: 0.7 µg/µL Normal Rabbit IgG. One vial containing 125 µg of Rabbit IgG in 125 µL of storage buffer containing 0.05% sodium azide. Store at -20°C. ChIP Primers, human β-globin. One vial containing 75 μL of 5 μM of each primer specific for the human β-globin promoter. Store at -20°C. FOR: AGG ACA GGT ACG GCT GTC ATC REV: TTT ATG CCC AGC CCT GGC TC
This ChIPAb+ Dimethyl-Histone H4 (Arg3) Symmetric -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications
Dot Blot
Western Blotting
Chromatin Immunoprecipitation (ChIP)
Application Notes
Chromatin Immunoprecipitation: Representative lot data. Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µg of either Normal rabbit IgG or 0.7 µg of Anti-Dimethyl-Histone H4 (Arg3) Symmetric and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of Dimethyl-Histone H4 (Arg3) Symmetric associated DNA fragments was verified by qPCR using ChIP Primers, human β-globin as a positive locus, and GAPDH promoter primers (22-004) as a negative locus. (Figure 2). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated. Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details. Western Blot Analysis: Representative lot data. HeLa acid extract (Lane 1) and recombinant Histone H4 (Lane 2) were probed with Anti-Dimethyl-Histone H4 (Arg3) Symmetrical (0.1μg/mL). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates Histone H4 (~13 kDa). (Figure 3). Note: An uncharacterized band at ~58 kDa has been observed in some lysates. Dot Blot Analysis: Representative lot data. Histone peptides with various modifications (see table below) were probed with Anti-Dimethyl-Histone H4 (Arg3) Symmetrical (1:2,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. (Figure 4). See table below. Immunocytochemistry Analysis: Representative lot data. Confocal fluorescent analysis of A431 and HeLa cells using Anti-Dimethyl-Histone H4 (Arg3) Symmetrical (Red). Actin filaments have been labeled with Alexa Fluor® 488 dye Phalloidin (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus. (Figure 5).
Biological Information
Immunogen
KLH-conjugated linear peptide corresponding to human Histone H4 when symmetrically dimethylated at Arg3.
Epitope
Dimethylated Arg3
Concentration
Please refer to the Certificate of Analysis for the lot-specific concentration.
Host
Rabbit
Specificity
This antibody recognizes Histone H4 symmetrically dimethylated at Arg3.
Species Reactivity
Human
Mouse
Rat
Species Reactivity Note
Demonstrated to react with human, mouse, and rat. Broad species cross-reactivity expected based on sequence similarity.
Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. This structure consists of approximately 146 bp of DNA wrapped around a nucleosome, an 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 H4 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is found in a histone cluster on chromosome 1. This gene is one of four histone genes in the cluster that are duplicated; this record represents the telomeric copy. [provided by RefSeq].
FUNCTION: 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. Chromosome.
PTM: Acetylation at Lys-6 (H4K5ac), Lys-9 (H4K8ac), Lys-13 (H4K12ac) and Lys-17 (H4K16ac) occurs in coding regions of the genome but not in heterochromatin.
Citrullination at Arg-4 (H4R3ci) by PADI4 impairs methylation.
Monomethylation at Arg-4 (H4R3me1) by PRMT1 favors acetylation at Lys-9 (H4K8ac) and Lys-13 (H4K12ac). Demethylation is performed by JMJD6.
Symmetric dimethylation on Arg-4 (H4R3me2a) by the PRDM1/PRMT5 complex may play a crucial role in the germ-cell lineage.
Monomethylated, dimethylated or trimethylated at Lys-21 (H4K20me1, H4K20me2, H4K20me3). Monomethylation is performed by SET8. Trimethylation is performed by SUV420H1 and SUV420H2 and induces gene silencing.
Ubiquitinated by the CUL4-DDB-RBX1 complex in response to ultraviolet irradiation. This may weaken the interaction between histones and DNA and facilitate DNA accessibility to repair proteins. Monoubiquitinated at Lys-92 of histone H4 (H4K91ub1) in response to DNA damage. The exact role of H4K91ub1 in DNA damage response is still unclear but it may function as a licensing signal for additional histone H4 post-translational modifications such as H4 Lys-21 methylation (H4K20me).
Sumoylated, which is associated with transcriptional repression.
SEQUENCE SIMILARITIES: Belongs to the histone H4 family.
SEQUENCE CAUTION: The sequence AAI28106.1 differs from that shown. Reason: Frameshift at position 3.
Molecular Weight
~13 kDa observed. Crossreacting band at ~58 kDa has been observed in some lysates.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
Chromatin Immunoprecipitation: Representative lot data. Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µg of either Normal Rabbit IgG, or 0.7 µg of Anti-Dimethyl-Histone H4 (Arg3) Symmetric and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of Dimethyl-Histone H4 (Arg3) Symmetric associated DNA fragments was verified by qPCR using ChIP Primers, human β-globin (Figure 1). Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
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 Conditions
Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
Packaging Information
Material Size
25 assays
Material Package
25 assays per set. Recommended use: 0.7 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Bestellnummer
GTIN
17-10250
04053252592348
Documentation
ChIPAb+™ Dimethyl-Histone H4 (Arg3) Symmetric - ChIP Validated Antibody and Primer Set SDB
Millipore’s Histone H4 antibodies demonstrate specificity against linker histone H4. See below for H4 Antibodies, ChipAb+ Primer sets, and peptides, based on the expertise of Upstate & Chemicon. Weitere Informationen >>
