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Epigenetics and Nuclear Signaling Histones Variants
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Anti-Histone H3.3 antibody (ab62642)

  • Datasheet
  • SDS
Reviews (7)Q&A (1)References (23)

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Western blot - Anti-Histone H3.3 antibody (ab62642)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3.3 antibody (ab62642)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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

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Secondary
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Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

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Overview

  • 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

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data
  • 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.
  • 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

  • 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

    • Epigenetics and Nuclear Signaling
    • Histones
    • Variants

Associated products

  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human Histone H3.3 protein (denatured) (ab156709)

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

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.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 19 kDa (predicted molecular weight: 15 kDa).

Target

  • 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
    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.
    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.
  • Target information above from: UniProt accession P84243 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    • Alternative names

      • H3 histone family 3A antibody
      • H3 histone family 3B antibody
      • H3 histone, family 3B (H3.3B) antibody
      • H3.3 antibody
      • H3.3A antibody
      • H3.3B antibody
      • H33_HUMAN antibody
      • H3F3 antibody
      • H3F3A antibody
      • H3f3b antibody
      • Histone H3.3 antibody
      • Histone H3.3Q antibody
      • Histone H3.A antibody
      • Histone H3.B antibody
      • MGC87782 antibody
      • MGC87783 antibody
      see all

    Images

    • Western blot - Anti-Histone H3.3 antibody (ab62642)
      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)
      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 (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

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (23)

    Publishing research using ab62642? Please let us know so that we can cite the reference in this datasheet.

    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
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-8 of 8 Abreviews or Q&A

    Western blot abreview for Anti-Histone H3.3 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293T cells cultured in DMEME+10% FBS)
    Gel Running Conditions
    Reduced Denaturing (15% precast Bis-Tris gel)
    Loading amount
    20 µg
    Treatment
    H3.3 with V5 tag over-expressed in pTRIEX 3 plasmid. Harvested 48h post-transfection
    Specification
    HEK293T cells cultured in DMEME+10% FBS
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 19 2017

    Western blot abreview for Anti-Histone H3.3 antibody

    Poor
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Zebrafish Cell lysate - whole cell (whole embryo, deyolked)
    Gel Running Conditions
    Reduced Denaturing (NuPAGE Bis-Tris gel 4-12%)
    Loading amount
    20000 cells
    Specification
    whole embryo, deyolked
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 24 2013

    Western blot abreview for Anti-Histone H3.3 antibody

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Raw 264.7)
    Gel Running Conditions
    Reduced Denaturing (12)
    Loading amount
    25 µg
    Treatment
    DRB and LPS 0-2hrs
    Specification
    Raw 264.7
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 31 2012

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3.3 antibody

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (osteosarcoma cell line, Saos2)
    Permeabilization
    Yes - PBS TRITON .25%
    Specification
    osteosarcoma cell line, Saos2
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 18 2012

    Question

    I would like to test this antibody in bull samples.

    Read More

    Abcam community

    Verified customer

    Asked on Feb 22 2012

    Answer

    To our knowledge, ab62642 has not been tested in bull. Therefore, I can offer a discount off a future purchase if you buy ab62642 now, test it inbull and submit feedback to us in the form of an Abreview. It doesn't matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value paid for the antibody.

    If you are interested in this offer, please let me know.

    Read More

    Abcam Scientific Support

    Answered on Feb 22 2012

    ChIP abreview for Anti-Histone H3.3 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Human Cell lysate - nuclear (Colon cancer cell)
    Negative control
    IgG control
    Specification
    Colon cancer cell
    Detection step
    Real-time PCR
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Read More

    Dr. santosh chauhan

    Verified customer

    Submitted Aug 02 2011

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3.3 antibody

    Poor
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Zebra finch Cell (HEK293 transfected with zebra finch H3.3 (GFP))
    Permeabilization
    Yes - 0.3% TritonX-100
    Specification
    HEK293 transfected with zebra finch H3.3 (GFP)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted May 27 2010

    ChIP abreview for Anti-Histone H3.3 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Human Cell lysate - whole cell (Soluble chromatin preparation from primary adipose)
    Negative control
    No antibody Dynabeads Protein A-only control
    Specification
    Soluble chromatin preparation from primary adipose
    Detection step
    Real-time PCR
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 8 minute(s) and 0 second(s)
    Specification of the cross-linking agent: Formaldehyde, 1%
    Positive control
    Promoter regions for which we have strong evidence of epitope tagged H3.3 enrichment in the human genome data provided byh ChIP-on-chip and ChIP-qPCR.
    Read More

    Prof. Philippe Collas

    Verified customer

    Submitted Oct 01 2009

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