Anti-Histone H3 antibody [mAbcam 24834] - Nuclear Loading Control and ChIP Grade (ab24834)
Key features and details
- Mouse monoclonal [mAbcam 24834] to Histone H3 - Nuclear Loading Control and ChIP Grade
- Suitable for: ChIP, WB
- Reacts with: Mouse, Rat, Human
- Isotype: IgG3
Overview
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Product name
Anti-Histone H3 antibody [mAbcam 24834] - Nuclear Loading Control and ChIP Grade
See all Histone H3 primary antibodies -
Description
Mouse monoclonal [mAbcam 24834] to Histone H3 - Nuclear Loading Control and ChIP Grade -
Host species
Mouse -
Tested Applications & Species
Application Species ChIP HumanWB MouseHuman -
Immunogen
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Positive control
- ChIP: Chromatin was prepared from HeLa cells. WB: HeLa histone prep. NIH/3T3 and PC-12 whole cell lysates. Mouse and rat testis tissue lysates.
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General notes
This antibody clone is manufactured by Abcam.
If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.50
Preservative: 0.02% Sodium azide
Constituent: PBS -
Concentration information loading...
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Purity
IgG fraction -
Clonality
Monoclonal -
Clone number
mAbcam 24834 -
Myeloma
Sp2/0-Ag14 -
Isotype
IgG3 -
Light chain type
kappa -
Research areas
Associated products
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Alternative Versions
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ChIP Related Products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab24834 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
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ChIP |
Human
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WB |
Mouse
Human
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All applications |
a wide range of other species
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Application | Abreviews | Notes |
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ChIP |
Use 5 µg for 25 µg of chromatin.
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WB | (6) |
Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 18 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 peptide (ab12149).
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Notes |
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ChIP
Use 5 µg for 25 µg of chromatin. |
WB
Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 18 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 peptide (ab12149). |
Target
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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
modificationsAcetylation 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. - Information by UniProt
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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
see all -
Alternative names
- H3 histone family member E pseudogene antibody
- H3 histone family, member A antibody
- H3/A antibody
see all
Images
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Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab24834 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.
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Western blot - Anti-Histone H3 antibody [mAbcam 24834] - Nuclear Loading Control and ChIP Grade (ab24834)Anti-Histone H3 antibody [mAbcam 24834] - Nuclear Loading Control and ChIP Grade (ab24834) at 1 µg/ml + HeLa histone prep at 1 µg/ml
Secondary
Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution
Predicted band size: 15 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted? -
Western blot - Anti-Histone H3 antibody [mAbcam 24834] - Nuclear Loading Control and ChIP Grade (ab24834)All lanes : Anti-Histone H3 antibody [mAbcam 24834] - Nuclear Loading Control and ChIP Grade (ab24834) at 1 µg/ml
Lane 1 : NIH/3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 2 : Testis (Mouse) Tissue Lysate
Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lane 4 : Testis (Rat) Tissue Lysate - normal tissue (ab29388)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutes
References (32)
ab24834 has been referenced in 32 publications.
- Laszig S et al. The novel dual BET/HDAC inhibitor TW09 mediates cell death by mitochondrial apoptosis in rhabdomyosarcoma cells. Cancer Lett 486:46-57 (2020). PubMed: 32445837
- Lobo-Silva J et al. The antischistosomal potential of GSK-J4, an H3K27 demethylase inhibitor: insights from molecular modeling, transcriptomics and in vitro assays. Parasit Vectors 13:140 (2020). PubMed: 32178714
- Pao PC et al. Examining the Role of HDACs in DNA Double-Strand Break Repair in Neurons. Methods Mol Biol 1983:225-234 (2019). PubMed: 31087301
- Currais A et al. Elevating acetyl-CoA levels reduces aspects of brain aging. Elife 8:N/A (2019). PubMed: 31742554
- Chung VY et al. The role of GRHL2 and epigenetic remodeling in epithelial-mesenchymal plasticity in ovarian cancer cells. Commun Biol 2:272 (2019). PubMed: 31372511