Anti-phospho (S10)-Acetyl (K14)-Histone H3 (rabbit polyclonal)

Species Reactivity	Key Applications	Host	Format	Antibody Type
M, H, Eu	ICC, WB, ChIP-seq, ChIP	Rb	Affinity Purified	Polyclonal Antibody

Species Reactivity

Key Applications

Host

Format

Antibody Type

M, H, Eu

ICC, WB, ChIP-seq, ChIP

Affinity Purified

Polyclonal Antibody

Description
Catalogue Number	07-081
Brand Family	Upstate
Trade Name	Upstate
Description	Anti-phospho (Ser10)-acetyl (Lys14)-Histone H3 Antibody
Alternate Names	H3K14me3S10P Histone H3 (acetyl K14, phospho S10) H3 histone family, member T histone 3 H3 histone cluster 3 H3
Background Information	Histone H3.1 (UniProt: Q16695; also known as H3K14AcS10P, Histone H3 (acetyl K14, phospho S10), H3 histone family member T, Histone 3, H3, Histone cluster 3, H3) is encoded by the HIST3H3 (also known as HIST1H3A, H3FA, HIST1H3B, H3FL, HIST1H3C, H3FC, HIST1H3D, H3FB, HIST1H3E, H3FD, HIST1H3F, H3FI, HIST1H3G, H3FH, HIST1H3H, H3FK, HIST1H3I, H3FF, HIST1H3J, H3FJ, HIST3H3, H3FT, H3T, MGC126886) gene (Gene ID: 8350, 8351, 8352, 8353, 8354, 8355, 8356, 8357, 8358, 8968) in human. Histone H3 has two main variants, H3.1 and H3.3, which show different genomic localization patterns in animals. The H3.1 and H3.3 complexes also possess distinct histone chaperones, CAF-1 and HIRA, which play important role in mediating DNA-synthesis-dependent and -independent nucleosome assembly. It has been reported that Histone H3.1 serves as the canonical histone, which is incorporated during DNA replication, whereas H3.3 acts as the replacement histone that can be incorporated outside of S-phase during chromatin-disrupting processes like transcription. Histone H 3.1 is a core component of nucleosome that is present only in mammals and is usually enriched in acetylation of Lysine 15 and demethylation of lysine 10 (HeK9Me2). It is expressed during S phase, then expression decreases significantly as cell division slows down during the process of differentiation. Histone H 3.1 expression is shown to be replication dependent. It s presence at the site of UV-induced DNA damage has also been reported. It has also been shown that H3.1/H4 tetramers do not split and remain intact during replication dependent deposition of H3.1 variant. Phosphorylation at serine 10 by aurora kinase B is reported to mediate the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin and this phosphorylation is considered to be an essential regulatory mechanism for neoplastic cell transformation. Acetylation of lysine 14 of histone H3 (H3K14ac) is shown to be critical for DNA damage checkpoint activation by directly regulating the compaction of chromatin and by recruiting chromatin remodeling protein complex RSC (Remodels the Structure of Chromatin). Nucleosomes with H3K14ac display higher affinity for purified RSC. (Ref.: Stroud, H., et al (2012). Proc. Natl. Acad. Sci. USA 109(14); 5370-75; Dunn, MR., and Smerdon MJ (2014). J. Biol. Chem. 289(12);8353-63)..

Description

Catalogue Number

07-081

Brand Family

Upstate

Trade Name

Upstate

Description

Anti-phospho (Ser10)-acetyl (Lys14)-Histone H3 Antibody

Alternate Names

H3K14me3S10P
Histone H3 (acetyl K14, phospho S10)
H3 histone family, member T
histone 3
H3
histone cluster 3
H3

Background Information

Histone H3.1 (UniProt: Q16695; also known as H3K14AcS10P, Histone H3 (acetyl K14, phospho S10), H3 histone family member T, Histone 3, H3, Histone cluster 3, H3) is encoded by the HIST3H3 (also known as HIST1H3A, H3FA, HIST1H3B, H3FL, HIST1H3C, H3FC, HIST1H3D, H3FB, HIST1H3E, H3FD, HIST1H3F, H3FI, HIST1H3G, H3FH, HIST1H3H, H3FK, HIST1H3I, H3FF, HIST1H3J, H3FJ, HIST3H3, H3FT, H3T, MGC126886) gene (Gene ID: 8350, 8351, 8352, 8353, 8354, 8355, 8356, 8357, 8358, 8968) in human. Histone H3 has two main variants, H3.1 and H3.3, which show different genomic localization patterns in animals. The H3.1 and H3.3 complexes also possess distinct histone chaperones, CAF-1 and HIRA, which play important role in mediating DNA-synthesis-dependent and -independent nucleosome assembly. It has been reported that Histone H3.1 serves as the canonical histone, which is incorporated during DNA replication, whereas H3.3 acts as the replacement histone that can be incorporated outside of S-phase during chromatin-disrupting processes like transcription. Histone H 3.1 is a core component of nucleosome that is present only in mammals and is usually enriched in acetylation of Lysine 15 and demethylation of lysine 10 (HeK9Me2). It is expressed during S phase, then expression decreases significantly as cell division slows down during the process of differentiation. Histone H 3.1 expression is shown to be replication dependent. It s presence at the site of UV-induced DNA damage has also been reported. It has also been shown that H3.1/H4 tetramers do not split and remain intact during replication dependent deposition of H3.1 variant. Phosphorylation at serine 10 by aurora kinase B is reported to mediate the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin and this phosphorylation is considered to be an essential regulatory mechanism for neoplastic cell transformation. Acetylation of lysine 14 of histone H3 (H3K14ac) is shown to be critical for DNA damage checkpoint activation by directly regulating the compaction of chromatin and by recruiting chromatin remodeling protein complex RSC (Remodels the Structure of Chromatin). Nucleosomes with H3K14ac display higher affinity for purified RSC. (Ref.: Stroud, H., et al (2012). Proc. Natl. Acad. Sci. USA 109(14); 5370-75; Dunn, MR., and Smerdon MJ (2014). J. Biol. Chem. 289(12);8353-63)..

Product Information
Format	Affinity Purified
Control	Acid-extracted proteins from serum-starved 10T1/2 cells.
Presentation	Purified rabbit polyclonal IgG in buffer containing 0.2 M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide, before the addition of glycerol to 30%.

Product Information

Format

Affinity Purified

Control

Acid-extracted proteins from serum-starved 10T1/2 cells.

Presentation

Purified rabbit polyclonal IgG in buffer containing 0.2 M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide, before the addition of glycerol to 30%.

Applications
Application	Anti-phospho (Ser10)-acetyl (Lys14)-Histone H3, Cat. No. 07-081, is a highly specific rabbit polyclonal antibody that targets Histone H3 phosphorylated on Serine 10 and acetylated on Lysine 14 and has been tested for use in Immunocytochemistry, Peptide Inhibition Assay, ChIP-seq and Chromatin Immuno
Key Applications	Immunocytochemistry Western Blotting ChIP-seq Chromatin Immunoprecipitation (ChIP)
Application Notes	Western Blot Analysis: 1:2000-1:10,000 dilutions of a previous lot detected phosphorylated and acetylated histone H3 in acid extracted proteins from serum starved mouse 10T1/2 cells treated with 50ng/ml EGF for 12.5 minutes. Western blotting of this lot to acid extracted proteins described above was preferentially competed by peptides in the following order: phospho-acetyl-H3 (7-20) > straight chain H3 (7-20) > phospho-H3 (7-20) or acetyl-H3 (7-20). Peptide Inhibition Analysis: A 1:500 dilution from a representative lot was used with HeLa AE for peptide block analysis. Chromatin Immunoprecipitation: A previous lot of this antibody was reported by an independent laboratory to immunoprecipitate chromatin. (Cheung, P., 2000.) Immunocytochemistry: A previous lot of this antibody detected phosphorylated and acetylated histone H3 in EGF-stimulated cells; as reported by an independent laboratory. (Cheung, P., 2000.)

Applications

Application

Anti-phospho (Ser10)-acetyl (Lys14)-Histone H3, Cat. No. 07-081, is a highly specific rabbit polyclonal antibody that targets Histone H3 phosphorylated on Serine 10 and acetylated on Lysine 14 and has been tested for use in Immunocytochemistry, Peptide Inhibition Assay, ChIP-seq and Chromatin Immuno

Key Applications

Immunocytochemistry
Western Blotting
ChIP-seq
Chromatin Immunoprecipitation (ChIP)

Application Notes

Western Blot Analysis:
1:2000-1:10,000 dilutions of a previous lot detected phosphorylated and acetylated histone H3 in acid extracted proteins from serum starved mouse 10T1/2 cells treated with 50ng/ml EGF for 12.5 minutes. Western blotting of this lot to acid extracted proteins described above was preferentially competed by peptides in the following order: phospho-acetyl-H3 (7-20) > straight chain H3 (7-20) > phospho-H3 (7-20) or acetyl-H3 (7-20).
Peptide Inhibition Analysis: A 1:500 dilution from a representative lot was used with HeLa AE for peptide block analysis.
Chromatin Immunoprecipitation:
A previous lot of this antibody was reported by an independent laboratory to immunoprecipitate chromatin. (Cheung, P., 2000.)

Immunocytochemistry:
A previous lot of this antibody detected phosphorylated and acetylated histone H3 in EGF-stimulated cells; as reported by an independent laboratory. (Cheung, P., 2000.)

Biological Information
Immunogen	KLH-conjugated, synthetic peptide corresponding to amino acids 7-20 of human histone H3 (ARK[pS]TGGAcKAPRKQL-C).
Concentration	Please refer to the Certificate of Analysis for the lot-specific concentration.
Host	Rabbit
Specificity	Recognizes Histone H3 phosphorylated at serine 10 and acetylated at lysine 14, Mr 17 kDa. An unidentified protein is also seen in some cells lines, Mr 40 kDa.
Isotype	IgG
Species Reactivity	Mouse Human Eukaryote
Species Reactivity Note	Human and mouse; broad species cross-reactivity expected.
Antibody Type	Polyclonal Antibody
Entrez Gene Number	NP_003484
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	HIST1H3A H3FA HIST1H3B H3FL HIST1H3C H3FC HIST1H3D H3FB HIST1H3E H3FD HIST1H3F H3FI HIST1H3G H3FH HIST1H3H H3FK HIST1H3I H3FF HIST1H3J H3FJ HIST3H3 H3FT H3T MGC126886
Modifications	Phosphorylation
Purification Method	Immunoaffinity and reverse-affinity chromatography.
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	17 kDa

Biological Information

Immunogen

KLH-conjugated, synthetic peptide corresponding to amino acids 7-20 of human histone H3 (ARK[pS]TGGAcKAPRKQL-C).

Concentration

Please refer to the Certificate of Analysis for the lot-specific concentration.

Host

Rabbit

Specificity

Recognizes Histone H3 phosphorylated at serine 10 and acetylated at lysine 14, Mr 17 kDa. An unidentified protein is also seen in some cells lines, Mr 40 kDa.

Isotype

IgG

Species Reactivity

Mouse
Human
Eukaryote

Species Reactivity Note

Human and mouse; broad species cross-reactivity expected.

Antibody Type

Polyclonal Antibody

Entrez Gene Number

NP_003484

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

HIST1H3A
H3FA
HIST1H3B
H3FL
HIST1H3C
H3FC
HIST1H3D
H3FB
HIST1H3E
H3FD
HIST1H3F
H3FI
HIST1H3G
H3FH
HIST1H3H
H3FK
HIST1H3I
H3FF
HIST1H3J
H3FJ
HIST3H3
H3FT
H3T
MGC126886

Modifications

Phosphorylation

Purification Method

Immunoaffinity and reverse-affinity chromatography.

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

17 kDa

Product Usage Statements
Quality Assurance	Routinely evaluated by Western Blot on HeLa acid extract. Western Blot Analysis: 1:500 dilution of this lot detected phospho acetyl Histone H3 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.

Product Usage Statements

Quality Assurance

Routinely evaluated by Western Blot on HeLa acid extract.

Western Blot Analysis:
1:500 dilution of this lot detected phospho acetyl Histone H3 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 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: Variabillity in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

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: Variabillity in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

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-phospho (S10)-Acetyl (K14)-Histone H3 (rabbit polyclonal)

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