Anti-dimethyl-Histone H3 (Arg2), clone 20.2 (rabbit monoclonal)

Species Reactivity	Key Applications	Host	Format	Antibody Type
H, Vrt	DB, Mplex, PIA, WB	Rb	Culture Supernatant	Monoclonal Antibody

Species Reactivity

Key Applications

Host

Format

Antibody Type

H, Vrt

DB, Mplex, PIA, WB

Culture Supernatant

Monoclonal Antibody

Description
Catalogue Number	04-808
Replaces	05-808
Brand Family	Upstate
Trade Name	Upstate
Description	Anti-dimethyl-Histone H3 (Arg2), clone 20.2 Antibody, rabbit monoclonal
Alternate Names	H3R2me2 Histone H3 (di methyl R2)
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). 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. The most commonly studied modifications are acetylation, phosphorylation, methylation, and ubiquitination. These modifications can alter local chromatin architecture, or recruit trans-acting factors that recognize specific histone modifications (the "histone code" hypothesis). The modifications occur predominantly on the N-terminal and C-terminal tails that extend beyond the nucleosome core particle. Histone H3 Arg2 methylation is a widespread silencing modification that inhibits histone H3 Lys4 trimethylation.

Description

Catalogue Number

04-808

Replaces

05-808

Brand Family

Upstate

Trade Name

Upstate

Description

Anti-dimethyl-Histone H3 (Arg2), clone 20.2 Antibody, rabbit monoclonal

Alternate Names

H3R2me2
Histone H3 (di methyl R2)

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). 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. The most commonly studied modifications are acetylation, phosphorylation, methylation, and ubiquitination. These modifications can alter local chromatin architecture, or recruit trans-acting factors that recognize specific histone modifications (the "histone code" hypothesis). The modifications occur predominantly on the N-terminal and C-terminal tails that extend beyond the nucleosome core particle. Histone H3 Arg2 methylation is a widespread silencing modification that inhibits histone H3 Lys4 trimethylation.

Product Information
Format	Culture Supernatant
Control	HeLa cell acid extracts
Presentation	Cultured supernantant in 0.05% sodium azide

Product Information

Format

Culture Supernatant

Control

HeLa cell acid extracts

Presentation

Cultured supernantant in 0.05% sodium azide

Applications
Application	Use Anti-dimethyl-Histone H3 (Arg2), clone 20.2 Antibody (rabbit monoclonal antibody) validated in DB, Mplex, PIA, WB to detect dimethyl-Histone H3 (Arg2) also known as H3R2me2, Histone H3 (di methyl R2).
Key Applications	Dot Blot Multiplexing Peptide Inhibition Assay Western Blotting
Application Notes	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. 04-808, Anti-dimethyl H3 (Arg2) at 1ug/mL (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 histone H3 peptide containing dimethyl-Arg2 abolished detection of histone H3 by a previous lot of anti-dimethyl-Histone H3(Arg2) (1:4,000 dilution) in immunoblots of HeLa acid extracts. Some competition of signal was also seen with unmodified peptide containing Arg2. Peptide Dot Blot Analysis: A 1:4,000-1:16,000 dilution of a previous lot only detected histone H3 peptide containing dimethyl-Arg2. Peptides containing unmodi-fied Arg2, dimethyl-Arg17 or dimethyl-Arg26 were not detected. Beadlyte® Histone Peptide Assay: 1:500-1:48,000 dilutions of a previous lot detected histone H3 peptide containing dimethyl-Arg2. Cross-reactivity to peptides containing unmodified Arg2 or dimethylArg 17 or 26 was not seen.

Applications

Application

Use Anti-dimethyl-Histone H3 (Arg2), clone 20.2 Antibody (rabbit monoclonal antibody) validated in DB, Mplex, PIA, WB to detect dimethyl-Histone H3 (Arg2) also known as H3R2me2, Histone H3 (di methyl R2).

Key Applications

Dot Blot
Multiplexing
Peptide Inhibition Assay
Western Blotting

Application Notes

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. 04-808, Anti-dimethyl H3 (Arg2) at 1ug/mL (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 histone H3 peptide containing dimethyl-Arg2 abolished detection of histone H3 by a previous lot of anti-dimethyl-Histone H3(Arg2) (1:4,000 dilution) in immunoblots of HeLa acid extracts. Some competition of signal was also seen with unmodified peptide containing Arg2.

Peptide Dot Blot Analysis:
A 1:4,000-1:16,000 dilution of a previous lot only detected histone H3 peptide containing dimethyl-Arg2. Peptides containing unmodi-fied Arg2, dimethyl-Arg17 or dimethyl-Arg26 were not detected.

Beadlyte® Histone Peptide Assay:
1:500-1:48,000 dilutions of a previous lot detected histone H3 peptide containing dimethyl-Arg2. Cross-reactivity to peptides containing unmodified Arg2 or dimethylArg 17 or 26 was not seen.

Biological Information
Immunogen	KLH-conjugated, synthetic peptide containing the sequence …Ame2RTKQ…, in which me2R corresponds to dimethyl-arginine 2 of human histone H3.
Epitope	Arg2 of Histone H3
Clone	20.2
Host	Rabbit
Specificity	Recognizes dimethyl-arginine at residue 2 of histone H3, Mr 17kDa. Additional unidentified bands above 50 kDa are detected in some samples.
Isotype	IgG
Species Reactivity	Human Vertebrates
Species Reactivity Note	Human. The immunizing sequence is highly conserved evolutionarily, so broad cross-reactivity is expected.
Antibody Type	Monoclonal Antibody
Entrez Gene Number	NM_003493
Entrez Gene Summary	Histones 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	HIST3H3 H3FT MGC126886 H3t MGC126888 H3T H3/g H3.4 H3/t
Modifications	Methylation
UniProt Number	Q16695
UniProt Summary	FUNCTION: SwissProt: Q16695 # 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: SwissProt: Q16695 ## Belongs to the histone H3 family.
Molecular Weight	17kDa

Biological Information

Immunogen

KLH-conjugated, synthetic peptide containing the sequence …Ame2RTKQ…, in which me2R corresponds to dimethyl-arginine 2 of human histone H3.

Epitope

Arg2 of Histone H3

Clone

20.2

Host

Rabbit

Specificity

Recognizes dimethyl-arginine at residue 2 of histone H3, Mr 17kDa. Additional unidentified bands above 50 kDa are detected in some samples.

Isotype

IgG

Species Reactivity

Human
Vertebrates

Species Reactivity Note

Human. The immunizing sequence is highly conserved evolutionarily, so broad cross-reactivity is expected.

Antibody Type

Monoclonal Antibody

Entrez Gene Number

NM_003493

Entrez Gene Summary

Histones 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

HIST3H3
H3FT
MGC126886
H3t
MGC126888
H3T
H3/g
H3.4
H3/t

Modifications

Methylation

UniProt Number

Q16695

UniProt Summary

FUNCTION: SwissProt: Q16695 # 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: SwissProt: Q16695 ## Belongs to the histone H3 family.

Molecular Weight

17kDa

Product Usage Statements
Quality Assurance	Routinely evaluated by immunoblot on acid-extracted proteins from HeLa cells. Immunoblot Analysis: A 1:500 (lane 1) and 1:1000 (lane 2) dilution of this lot detected methylated Histone H3 in acid-extracted proteins from HeLa cells.
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.

Product Usage Statements

Quality Assurance

Routinely evaluated by immunoblot on acid-extracted proteins from HeLa cells.

Immunoblot Analysis: A 1:500 (lane 1) and 1:1000 (lane 2) dilution of this lot detected methylated Histone H3 in acid-extracted proteins from HeLa cells.

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 receipt, 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.

Storage and Shipping Information

Storage Conditions

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt, 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.

Packaging Information
Material Size	200 µL

Packaging Information

Material Size

200 µL

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

办公用品

Anti-dimethyl-Histone H3 (Arg2), clone 20.2 (rabbit monoclonal)

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