Name: Anti-Histone H3 (phospho S10 + T11) antibody [E173]
Brand: RabMAb
SKU: ab32107
Rating: 5 (4 reviews)

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Western blot - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [E173] to Histone H3 (phospho S10 + T11)
Suitable for: ELISA, Dot blot, ICC/IF, WB, IHC-P, IP
Reacts with: Mouse, Human

Alexa Fluor® 488 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free HRP

Product name

Anti-Histone H3 (phospho S10 + T11) antibody [E173]
See all Histone H3 primary antibodies
Description

Rabbit monoclonal [E173] to Histone H3 (phospho S10 + T11)
Host species

Rabbit
Specificity

This antibody detects Histone H3 phosphorylated on both Serine 10 and Threonine 11. However, the antibody shows higher affinity for phosphorylated Serine 10 than for phosphorylated Threonine 11. This was validated by ELISA, Dot Blot and WB peptide blocking experiments.
Tested applications

Suitable for: ELISA, Dot blot, ICC/IF, WB, IHC-P, IPmore details
Unsuitable for: Flow Cyt
Species reactivity

Reacts with: Mouse, Human
Predicted to work with: Rat, Drosophila melanogaster, a wide range of other species
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HeLa whole cell lysate (ab150035) treated with Calyculin A. IHC-P: Lymphoma tissue. ICC/IF: HeLa cells IP: HeLa treated with calyculin A whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

E173
Isotype

IgG
Research areas

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32107 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
ELISA		Use at an assay dependent concentration.
Dot blot		1/1000.
ICC/IF	(1)	1/250 - 1/1000.
WB	(1)	1/2500. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).
IHC-P	(2)	Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP		1/50.

Notes
ELISA Use at an assay dependent concentration.
Dot blot 1/1000.
ICC/IF 1/250 - 1/1000.
WB 1/2500. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP 1/50.

Application notes

Is unsuitable for Flow Cyt.

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.
Sequence similarities

Belongs to the histone H3 family.
Developmental stage

Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
Post-translational
modifications

Acetylation is generally linked to gene activation. Acetylation on Lys-10 (H3K9ac) impairs methylation at Arg-9 (H3R8me2s). Acetylation on Lys-19 (H3K18ac) and Lys-24 (H3K24ac) favors methylation at Arg-18 (H3R17me).
Citrullination at Arg-9 (H3R8ci) and/or Arg-18 (H3R17ci) by PADI4 impairs methylation and represses transcription.
Asymmetric dimethylation at Arg-18 (H3R17me2a) by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 (H3R8me2s) by PRMT5 is linked to gene repression. Asymmetric dimethylation at Arg-3 (H3R2me2a) by PRMT6 is linked to gene repression and is mutually exclusive with H3 Lys-5 methylation (H3K4me2 and H3K4me3). H3R2me2a is present at the 3' of genes regardless of their transcription state and is enriched on inactive promoters, while it is absent on active promoters.
Methylation at Lys-5 (H3K4me), Lys-37 (H3K36me) and Lys-80 (H3K79me) are linked to gene activation. Methylation at Lys-5 (H3K4me) facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 (H3K79me) is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are linked to gene repression. Methylation at Lys-10 (H3K9me) is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 (H3S10ph) and acetylation of H3 and H4. Methylation at Lys-5 (H3K4me) and Lys-80 (H3K79me) require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are enriched in inactive X chromosome chromatin.
Phosphorylated at Thr-4 (H3T3ph) by GSG2/haspin during prophase and dephosphorylated during anaphase. Phosphorylation at Ser-11 (H3S10ph) by AURKB is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at Ser-11 (H3S10ph) by RPS6KA4 and RPS6KA5 is important during interphase because it enables the transcription of genes following external stimulation, like mitogens, stress, growth factors or UV irradiation and result in the activation of genes, such as c-fos and c-jun. Phosphorylation at Ser-11 (H3S10ph), which is linked to gene activation, prevents methylation at Lys-10 (H3K9me) but facilitates acetylation of H3 and H4. Phosphorylation at Ser-11 (H3S10ph) by AURKB mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. Phosphorylation at Ser-11 (H3S10ph) is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylated at Ser-29 (H3S28ph) by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation at Thr-7 (H3T6ph) by PRKCBB is a specific tag for epigenetic transcriptional activation that prevents demethylation of Lys-5 (H3K4me) by LSD1/KDM1A. At centromeres, specifically phosphorylated at Thr-12 (H3T11ph) from prophase to early anaphase, by DAPK3 and PKN1. Phosphorylation at Thr-12 (H3T11ph) by PKN1 is a specific tag for epigenetic transcriptional activation that promotes demethylation of Lys-10 (H3K9me) by KDM4C/JMJD2C. Phosphorylation at Tyr-42 (H3Y41ph) by JAK2 promotes exclusion of CBX5 (HP1 alpha) from chromatin.
Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination (By similarity). 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.
Cellular localization

Nucleus. Chromosome.
Target information above from: UniProt accession P68431 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 8350 Human
- Entrez Gene: 8351 Human
- Entrez Gene: 8352 Human
- Entrez Gene: 8353 Human
- Entrez Gene: 8354 Human
- Entrez Gene: 8355 Human
- Entrez Gene: 8356 Human
- Entrez Gene: 8357 Human
- Entrez Gene: 8358 Human
- Entrez Gene: 8968 Human
- Entrez Gene: 319152 Mouse
- Entrez Gene: 319153 Mouse
- Entrez Gene: 360198 Mouse
- Entrez Gene: 97908 Mouse
- Entrez Gene: 100364501 Rat
- Entrez Gene: 100365669 Rat
- Entrez Gene: 291159 Rat
- Entrez Gene: 679994 Rat
- Entrez Gene: 680511 Rat
- Entrez Gene: 682330 Rat
- Omim: 602810 Human
- SwissProt: P68431 Human
- SwissProt: P68433 Mouse
- SwissProt: Q6LED0 Rat
- Unigene: 132854 Human
- Unigene: 247813 Human
- Unigene: 247814 Human
- Unigene: 248176 Human
- Unigene: 443021 Human
- Unigene: 484990 Human
- Unigene: 532144 Human
- Unigene: 533292 Human
- Unigene: 546315 Human
- Unigene: 586261 Human
- Unigene: 591778 Human
- Unigene: 221301 Mouse
- Unigene: 261657 Mouse
- Unigene: 377874 Mouse
- Unigene: 390558 Mouse
- Unigene: 397328 Mouse
- Unigene: 138090 Rat
see all
Alternative names
- H3 histone family member E pseudogene antibody
- H3 histone family, member A antibody
- H3/A antibody
- H31_HUMAN antibody
- H3F3 antibody
- H3FA antibody
- Hist1h3a antibody
- HIST1H3B antibody
- HIST1H3C antibody
- HIST1H3D antibody
- HIST1H3E antibody
- HIST1H3F antibody
- HIST1H3G antibody
- HIST1H3H antibody
- HIST1H3I antibody
- HIST1H3J antibody
- HIST3H3 antibody
- histone 1, H3a antibody
- Histone cluster 1, H3a antibody
- Histone H3 3 pseudogene antibody
- Histone H3.1 antibody
- Histone H3/a antibody
- Histone H3/b antibody
- Histone H3/c antibody
- Histone H3/d antibody
- Histone H3/f antibody
- Histone H3/h antibody
- Histone H3/i antibody
- Histone H3/j antibody
- Histone H3/k antibody
- Histone H3/l antibody
see all

Western blot - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

All lanes : Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107) at 1/10000 dilution

Lane 1 : Untreated HeLa cell lysate
Lane 2 : HeLa cell lysate treated with calyculin A
Lane 3 : HeLa cell lysate treated with calyculin A and alkaline phosphatase

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 15 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?

Exposure time: 1 second

Blocking and dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling Histone H3 (phospho S10 + T11) with ab32107 at a dilution of 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000, 2 μg/mL) was used as the secondary antibody. Cells were counter-stained with ab195889 Anti-Alpha Tubulin antibody [DM1A] (1/200, 2.5 μg/mL) - Microtubule Marker (Alexa Fluor^® 594). DAPI (blue) was used as a nuclear counterstain.
Confocal image showing nuclear staining on mitotic HeLa cell.
Immunoprecipitation - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

Purified ab32107 at 1/50 dilution (2µg) immunoprecipitating Histone H3 in HeLa treated with calyculin A whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) treated with calyculin A whole cell lysate 10µg
Lane 2 (+): ab32107 + HeLa treated with calyculin A whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32107 in HeLa treated with calyculin A whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 17 kDa
Western blot - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

All lanes : Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107) at 1/1000 dilution

Lane 1 : HeLa whole cell lysate -treated with Calyculin A with Histone H3 phospho S10 peptide
Lane 2 : HeLa whole cell lysate -treated with Calyculin A with Histone H3 phospho T11 peptide
Lane 3 : HeLa whole cell lysate -treated with Calyculin A with unmodified Histone H3 peptide
Lane 4 : HeLa whole cell lysate -treated with Calyculin A with no peptide

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

Predicted band size: 15 kDa
Observed band size: 15 kDa

Exposure time: 1 second

Blocking/Diluting buffer 5% NFDM/TBST
Dot Blot - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

Dot blot analysis of Histone H3 phospho S10 peptide (Lane 1), Histone H3 phospho T11 peptide (Lane 2) and Histone H3 unmodified peptide (Lane 3) labelling Histone H3 (phospho S10 + T11) with ab32107 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG) (H+L) was used as the secondary antibody at a dilution of 1/10 000.

Blocking and diluting buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

Ab32107, at a 1/50 dilution, staining Histone H3 (phospho S10 + T11) in paraffin embedded lymphoma tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)This image is courtesy of an Abreview submitted by Dr Kirk McManus

ab32107 (1/1000) staining Histone H3 (phospho S10 + T11) in paraformaldehyde-fixed, DAPI counterstained HeLa cells. Secondary antibody: Goat anti-Rabbit IgG conjugated to Cy3® (1/200). Please refer to abreview for further details.
See Abreview
ELISA - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

Direct ELISA antibody dose-response curve using ab32107 (0-1000 ng/mL).

Peptides - Histone H3 unmodified peptide, Histone H3 phospho S10 peptide, Histone H3 phospho T11 peptide (100 ng/mL).

Secondary antibody - Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (1/2500).
ELISA - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

Direct ELISA antibody dose-response curve using ab32107 (0-1000 ng/mL).

Peptides - Histone H3 unmodified peptide, Histone H3 phospho S10 peptide, Histone H3 phospho T11 peptide (10 ng/mL).

Secondary antibody - Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (1/2500).
Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab32107? Please let us know so that we can cite the reference in this datasheet.

ab32107 has been referenced in 42 publications.

Wu Y et al. LRP6 downregulation promotes cardiomyocyte proliferation and heart regeneration. Cell Res 31:450-462 (2021). PubMed: 32973339
Niu K et al. Canonical and early lineage-specific stem cell types identified in planarian SirNeoblasts. Cell Regen 10:15 (2021). PubMed: 33740162
Bohr TE et al. Planarian stem cells sense the identity of the missing pharynx to launch its targeted regeneration. Elife 10:N/A (2021). PubMed: 34156924
Shiroor DA et al. Injury Delays Stem Cell Apoptosis after Radiation in Planarians. Curr Biol 30:2166-2174.e3 (2020). PubMed: 32386527
Lin TY et al. BIRC5/Survivin is a novel ATG12-ATG5 conjugate interactor and an autophagy-induced DNA damage suppressor in human cancer and mouse embryonic fibroblast cells. Autophagy 16:1296-1313 (2020). PubMed: 31612776

View all Publications for this product

Customer reviews and Q&As

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1-4 of 4 Abreviews or Q&A

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Cat Tissue sections (Skin tumor)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: BH6 citrate buffer

Permeabilization

Specification

Skin tumor

Blocking step

BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 24°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 24 2021

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Dog Tissue sections (Skin, tumor)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: citrate buffer

Permeabilization

Specification

Skin, tumor

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 20% · Temperature: 20°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 01 2020

Application

Western blot

Loading amount

30 µg

Gel Running Conditions

Reduced Denaturing (10%)

Sample

Fruit fly (Drosophila melanogaster) Tissue lysate - whole (0-4hr embryos)

Specification

0-4hr embryos

Treatment

control shRNA and wee shRNA embryos

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Richelle Sopko

Verified customer

Submitted Dec 04 2013

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (HeLa)

Specification

HeLa

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Kirk Mcmanus

Verified customer

Submitted Feb 20 2007

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Developmental stage

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (42)

Customer reviews and Q&As

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Histone H3 (phospho S10 + T11) antibody [E173]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Histone H3 (phospho S10 + T11) antibody [E173]

Western blot abreview for Anti-Histone H3 (phospho S10) antibody [E173]

Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (phospho S10) antibody [E173]

Post-translational
modifications