Anti-dimethyl-Histone H3 (Lys79), clone NL59 (rabbit monoclonal)

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

Species Reactivity

Key Applications

Host

Format

Antibody Type

H, Vrt

WB, DB, Mplex, ChIP-seq, ChIP

Culture Supernatant

Monoclonal Antibody

Description
Catalogue Number	04-835
Replaces	05-835
Brand Family	Upstate
Trade Name	Upstate
Description	Anti-dimethyl-Histone H3 (Lys79) Antibody, clone NL59, rabbit monoclonal
Alternate Names	H3K79me2 Histone H3 (di methyl K79)
Background Information	Histone 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 Lys79 on histone H3 is associated with active gene transcription and is a marker for euchromatin. Histone H3 is methylated on Lys79 by the histone methyltransferase Dot1L..

Description

Catalogue Number

04-835

Replaces

05-835

Brand Family

Upstate

Trade Name

Upstate

Description

Anti-dimethyl-Histone H3 (Lys79) Antibody, clone NL59, rabbit monoclonal

Alternate Names

H3K79me2
Histone H3 (di methyl K79)

Background Information

Histone 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 Lys79 on histone H3 is associated with active gene transcription and is a marker for euchromatin. Histone H3 is methylated on Lys79 by the histone methyltransferase Dot1L..

Product Information
Format	Culture Supernatant
Presentation	Culture supernantant in 0.05% sodium azide

Product Information

Format

Culture Supernatant

Presentation

Culture supernantant in 0.05% sodium azide

Applications
Application	Anti-dimethyl-Histone H3 (Lys79) Antibody, clone NL59 is a rabbit monoclonal antibody for detection of dimethyl-Histone H3 (Lys79) also known as H3K79me2, Histone H3 (di methyl K79) & has been validated in ChIP, WB, DB, Mplex.
Key Applications	Western Blotting Dot Blot Multiplexing ChIP-seq Chromatin Immunoprecipitation (ChIP)
Application Notes	Chromatin Immunoprecipitation: Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 4 µL of Negative Control Supernatant, or 4 µL of Anti-dimethyl-Histone H3 (Lys79) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of dimethyl-Histone H3 (Lys79) associated DNA fragments was verified by qPCR using Control Primers Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details. Chromatin Immunoprecipitation: A representative lot of this antibody has been shown by an independent laboratory to work in ChIP. ChIP-Seq Analysis: A representative lot of this antibody was used by an independent laboratory for ChIP-Seq. See Egelhofer, T.A., et al. (2011). See Easwaran, H., et al. (2012).See Suzuki, H., et al. (2011). Dot Blotting: Specificity of a representative lot confirmed by the ability of a 1:2500 dilution of the antibody to recognize peptides corresponding to regions of histone H3 with various modifications 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 04-835-S, Anti-dimethyl Histone H3 (Lys79) Antibody, clone NL59 (1:500 dilution). Proteins were visualized using a Donkey anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system.

Applications

Application

Anti-dimethyl-Histone H3 (Lys79) Antibody, clone NL59 is a rabbit monoclonal antibody for detection of dimethyl-Histone H3 (Lys79) also known as H3K79me2, Histone H3 (di methyl K79) & has been validated in ChIP, WB, DB, Mplex.

Key Applications

Western Blotting
Dot Blot
Multiplexing
ChIP-seq
Chromatin Immunoprecipitation (ChIP)

Application Notes

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 4 µL of Negative Control Supernatant, or 4 µL of Anti-dimethyl-Histone H3 (Lys79) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of dimethyl-Histone H3 (Lys79) associated DNA fragments was verified by qPCR using Control Primers
Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

Chromatin Immunoprecipitation: A representative lot of this antibody has been shown by an independent laboratory to work in ChIP.

ChIP-Seq Analysis:
A representative lot of this antibody was used by an independent laboratory for ChIP-Seq. See Egelhofer, T.A., et al. (2011). See Easwaran, H., et al. (2012).See Suzuki, H., et al. (2011).

Dot Blotting: Specificity of a representative lot confirmed by the ability of a 1:2500 dilution of the antibody to recognize peptides corresponding to regions of histone H3 with various modifications

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 04-835-S, Anti-dimethyl Histone H3 (Lys79) Antibody, clone NL59 (1:500 dilution). Proteins were visualized using a Donkey anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system.

Biological Information
Immunogen	KLH-conjugated synthetic peptide containing the sequence DF_me2KTD corresponding to dimethyl-lysine at position 79 of histone H3. The immunizing sequence is identical in human, mouse, rat and yeast. A C-terminal cysteine was added to facilitate conjugation.
Epitope	Lys79
Clone	NL59
Host	Rabbit
Specificity	Recognizes dimethyl-histone H3 (Lys79).
Isotype	IgG
Species Reactivity	Human Vertebrates
Species Reactivity Note	Broad species cross-reactivity expected due to sequence homology
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.
Gene Symbol	H3F3A H3F3B
Modifications	Methylation
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	17 kDa

Biological Information

Immunogen

KLH-conjugated synthetic peptide containing the sequence DF_me2KTD corresponding to dimethyl-lysine at position 79 of histone H3. The immunizing sequence is identical in human, mouse, rat and yeast. A C-terminal cysteine was added to facilitate conjugation.

Epitope

Lys79

Clone

NL59

Host

Rabbit

Specificity

Recognizes dimethyl-histone H3 (Lys79).

Isotype

IgG

Species Reactivity

Human
Vertebrates

Species Reactivity Note

Broad species cross-reactivity expected due to sequence homology

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.

Gene Symbol

H3F3A
H3F3B

Modifications

Methylation

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

17 kDa

Product Usage Statements
Quality Assurance	routinely evaluated by immunoblot of 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 of 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	100 µL

Packaging Information

Material Size

100 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 (Lys79), clone NL59 (rabbit monoclonal)

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