Anti-Histone H3.3 antibody (ab62642)
Key features and details
- Rabbit polyclonal to Histone H3.3
- Suitable for: ICC/IF, IHC-P, WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-Histone H3.3 antibody
See all Histone H3.3 primary antibodies -
Description
Rabbit polyclonal to Histone H3.3 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanIHC-P HumanWB Human
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Immunogen
Synthetic peptide corresponding to Human Histone H3.3. (synthesized peptide derived from Internal of human Histone H3.3.)
Database link: P84243 -
Positive control
- WB: A431 (Human epithelial carcinoma cell line). ICC: HeLa cells. IHC-P: Human brain tissue.
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
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. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7
Preservative: 0.02% Sodium azide
Constituents: PBS, 50% Glycerol (glycerin, glycerine), 0.87% Sodium chloride -
Concentration information loading... -
Purity
Immunogen affinity purified -
Purification notes
The antibody was affinity purified from rabbit antiserum by affinity chromatography using epitope specific immunogen. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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ChIP Related Products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab62642 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 |
|---|---|
| ICC/IF |
Human
|
| IHC-P |
Human
|
| WB |
Human
|
| Application | Abreviews | Notes |
|---|---|---|
| ICC/IF | (2) |
1/500 - 1/1000.
|
| IHC-P |
1/50 - 1/100.
|
|
| WB | (3) |
Use a concentration of 1 µg/ml. Detects a band of approximately 19 kDa (predicted molecular weight: 15 kDa).
|
| Notes |
|---|
|
ICC/IF
1/500 - 1/1000. |
|
IHC-P
1/50 - 1/100. |
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 19 kDa (predicted molecular weight: 15 kDa). |
Target
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Function
Variant histone H3 which replaces conventional H3 in a wide range of nucleosomes in active genes. Constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. Deposited at sites of nucleosomal displacement throughout transcribed genes, suggesting that it represents an epigenetic imprint of transcriptionally active chromatin. 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 throughout the cell cycle independently of DNA synthesis. -
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.
Specifically enriched in modifications associated with active chromatin such as methylation at Lys-5 (H3K4me), Lys-37 and Lys-80. 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), which are linked to gene repression, are underrepresented. 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. Phosphorylation on Ser-32 (H3S31ph) is specific to regions bordering centromeres in metaphase chromosomes.
Ubiquitinated. Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination. -
Cellular localization
Nucleus. Chromosome. - Information by UniProt
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Database links
- Entrez Gene: 3020 Human
- Entrez Gene: 3021 Human
- Omim: 601058 Human
- SwissProt: P84243 Human
- Unigene: 180877 Human
- Unigene: 533624 Human
- Unigene: 726012 Human
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Alternative names
- H3 histone family 3A antibody
- H3 histone family 3B antibody
- H3 histone, family 3B (H3.3B) antibody
see all
Images
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Western blot - Anti-Histone H3.3 antibody (ab62642)Anti-Histone H3.3 antibody (ab62642) at 1 µg/ml + A431 (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 19 kDa why is the actual band size different from the predicted?
Additional bands at: 45 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 12 minutes
The 19 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to Histone H3.3. -
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3.3 antibody (ab62642)Immunofluorescence analysis of HeLa cells, using ab62642 at a 1/500 dilution.
Left image untreated.
Right image treated with peptide. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 antibody (ab62642)Immunohistochemistry analysis of paraffin embedded human brain tissue using ab62642 at a 1/50 dilution.
Left image untreated.
Right image treated with peptide.
Protocols
References (23)
ab62642 has been referenced in 23 publications.
- Zhu Y et al. Partial hepatectomy-induced upregulation of SNHG12 promotes hepatocyte proliferation and liver regeneration. Mol Med Rep 21:1089-1096 (2020). PubMed: 31894329
- Coulombe P et al. The ORC ubiquitin ligase OBI1 promotes DNA replication origin firing. Nat Commun 10:2426 (2019). PubMed: 31160578
- Bagati A et al. KLF9-dependent ROS regulate melanoma progression in stage-specific manner. Oncogene 38:3585-3597 (2019). PubMed: 30664687
- Rachdaoui N et al. Turnover of histones and histone variants in postnatal rat brain: effects of alcohol exposure. Clin Epigenetics 9:117 (2017). PubMed: 29075360
- Bassat E et al. The extracellular matrix protein agrin promotes heart regeneration in mice. Nature 547:179-184 (2017). PubMed: 28581497
