Anti-phospho-Histone H3 (Thr11), clone MC83 (rabbit monoclonal)

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

Species Reactivity

Key Applications

Host

Format

Antibody Type

WB, PIA, Mplex

Culture Supernatant

Monoclonal Antibody

Description
Catalogue Number	04-789
Replaces	MABE468
Brand Family	Upstate
Trade Name	Upstate
Description	Anti-phospho-Histone H3 (Thr11) Antibody, clone MC83, rabbit monoclonal
Alternate Names	H3T11P Histone H3 (phospho T11)
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. Phosphorylation at Ser10, Ser28 and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis. In mammals, phosphorylation of Thr11 modification is restricted to the centromeric region while in plant cells, phosphorylation of Thr11 was distributed along the entire length of condensed chromosomes. Mitosis-specific phosphorylation of histone H3 at Thr11 is mediated by Dlk/ZIP kinase.

Description

Catalogue Number

04-789

Replaces

MABE468

Brand Family

Upstate

Trade Name

Upstate

Description

Anti-phospho-Histone H3 (Thr11) Antibody, clone MC83, rabbit monoclonal

Alternate Names

H3T11P
Histone H3 (phospho T11)

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. Phosphorylation at Ser10, Ser28 and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis. In mammals, phosphorylation of Thr11 modification is restricted to the centromeric region while in plant cells, phosphorylation of Thr11 was distributed along the entire length of condensed chromosomes. Mitosis-specific phosphorylation of histone H3 at Thr11 is mediated by Dlk/ZIP kinase.

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

Product Information

Format

Culture Supernatant

Control

HeLa acid extracts

Presentation

Cultured supernantant in 0.05% sodium azide

Applications
Application	Anti-phospho-Histone H3 (Thr11) Antibody, clone MC83 is a rabbit monoclonal antibody for detection of phospho-Histone H3 (Thr11) also known as H3T11P, Histone H3 (phospho T11) & has been validated in WB, PIA, Mplex.
Key Applications	Western Blotting Peptide Inhibition Assay Multiplexing
Application Notes	Immunoblot Analysis: A 1:500-1:2000 dilution of this lot detected phosphorylated histone H3 in acid extracted proteins from mitotic HeLa cells treated with colcemid, but not unmodified recombinant histone H3 (Catalog # 14-494). Western Blot Analysis: A 1:5,000 dilution of a previous lot detected phosphorylated histone H3 in acid extracted proteins from mitotic HeLa cells treated with colcemid, but not unmodified recombinant histone H3 (Catalog # 14-494). Multiplexing: Beadlyte® Histone-Peptide Specificity Assay: 1:1,000-1:50,000 dilutions of a previous lot were incubated with histone H3 peptides containing various modifications conjugated to Luminex® microspheres. Only the peptide containing phospho-threonine 11 was detected (Figure C). Peptide Inhibition Analysis: 1 μM of histone H3 peptide containing phospho-threonine 11 abolished detection of histone H3 by anti-phospho-Histone H3 (Thr11), clone MC83 in immunoblot analysis of acid extracts from colcemid treated HeLa cells (Figure D, lane 4).

Applications

Application

Anti-phospho-Histone H3 (Thr11) Antibody, clone MC83 is a rabbit monoclonal antibody for detection of phospho-Histone H3 (Thr11) also known as H3T11P, Histone H3 (phospho T11) & has been validated in WB, PIA, Mplex.

Key Applications

Western Blotting
Peptide Inhibition Assay
Multiplexing

Application Notes

Immunoblot Analysis:
A 1:500-1:2000 dilution of this lot detected phosphorylated histone H3 in acid extracted proteins from mitotic HeLa cells treated with colcemid, but not unmodified recombinant histone H3 (Catalog # 14-494).

Western Blot Analysis:
A 1:5,000 dilution of a previous lot detected phosphorylated histone H3 in acid extracted proteins from mitotic HeLa cells treated with colcemid, but not unmodified recombinant histone H3 (Catalog # 14-494).

Multiplexing:
Beadlyte® Histone-Peptide Specificity Assay: 1:1,000-1:50,000 dilutions of a previous lot were incubated with histone H3 peptides containing various modifications conjugated to Luminex® microspheres. Only the peptide containing phospho-threonine 11 was
detected (Figure C).

Peptide Inhibition Analysis: 1 μM of histone H3 peptide containing phospho-threonine 11 abolished detection of histone H3 by anti-phospho-Histone H3 (Thr11), clone MC83 in immunoblot analysis of acid extracts from
colcemid treated HeLa cells (Figure D, lane 4).

Biological Information
Immunogen	Peptide containing ...KS[pT]G..., in which pT corresponds to phospho-threonine at position 11 of human histone H3
Clone	MC83
Host	Rabbit
Specificity	Recognizes histone H3 phosphorylated at Thr11, Mr 17kDa.
Isotype	IgG
Species Reactivity	Human
Species Reactivity Note	Human. Broad species cross-reactivity is expected based on sequence homology. An additional unidentified band at ~40 kDa appears in some lysates.
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	H3/t H3.4 H3/g H3T MGC126888 H3t MGC126886 H3FT HIST3H3
Modifications	Phosphorylation
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

Peptide containing ...KS[pT]G..., in which pT corresponds to phospho-threonine at position 11 of human histone H3

Clone

MC83

Host

Rabbit

Specificity

Recognizes histone H3 phosphorylated at Thr11, Mr 17kDa.

Isotype

IgG

Species Reactivity

Human

Species Reactivity Note

Human. Broad species cross-reactivity is expected based on sequence homology. An additional unidentified band at ~40 kDa appears in some lysates.

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

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

Modifications

Phosphorylation

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 analysis.
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 analysis.

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. For maximum recovery of product, centrifuge the vial prior to removing the cap.

Storage and Shipping Information

Storage Conditions

Stable for 1 year at -20°C from date of receipt.
For maximum recovery of product, centrifuge the vial prior to removing the cap.

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-phospho-Histone H3 (Thr11), clone MC83 (rabbit monoclonal)

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