RecombinantRabMAb

Recombinant Alexa Fluor® 647 Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab237478)

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Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab237478)
  • Flow Cytometry - Alexa Fluor® 647 Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab237478)
  • Alexa Fluor® 647 Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab237478)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Alexa Fluor® 647 Rabbit monoclonal [EPR1873] to Histone H3.3 (phospho S31)
  • Suitable for: Flow Cyt, ICC/IF
  • Reacts with: Human
  • Conjugation: Alexa Fluor® 647. Ex: 652nm, Em: 668nm

Overview

  • Product name

    Alexa Fluor® 647 Anti-Histone H3.3 (phospho S31) antibody [EPR1873]
    See all Histone H3.3 primary antibodies

  • Description

    Alexa Fluor® 647 Rabbit monoclonal [EPR1873] to Histone H3.3 (phospho S31)

  • Host species

    Rabbit

  • Conjugation

    Alexa Fluor® 647. Ex: 652nm, Em: 668nm

  • Specificity

    Detects Histone H3.3 when phosphorylated at S31.

  • Tested applications

    Suitable for: Flow Cyt, ICC/IFmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat

  • Immunogen

    Synthetic peptide within Human Histone H3.3 aa 1-100 (phospho S31). The exact sequence is proprietary.
    Database link: P84243

  • Positive control

    • Flow Cyt: HeLa cells. ICC/IF: HeLa cells

  • 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.

    Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

    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. Stable for 12 months at -20°C. Store In the Dark.

  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: 30% Glycerol, 1% BSA, PBS
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR1873

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab237478 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
Flow Cyt 1/500.
ICC/IF 1/100.

This product gave a positive signal in HeLa cells fixed with 4% formaldehyde (10 min)

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.
  • Information by UniProt

  • Database links

    see all

  • 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

  • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab237478)
    Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab237478)

    ab237478 staining Histone H3.3 (phospho S31) in HeLa cells +/- Lambda Phosphatase (2 hours at 31oC). The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h.

    The cells were then incubated overnight at +4°C with ab237478 at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Flow Cytometry - Alexa Fluor® 647 Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab237478)
    Flow Cytometry - Alexa Fluor® 647 Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab237478)

    Overlay histogram showing HeLa cells stained with ab237478 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 90% methanol. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab237478, 1/500 dilution) for 30 min at 22°C.

    Isotype control antibody (black line) was Rabbit IgG (monoclonal) Alexa Fluor® 647 (ab199093) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 17mW red Helium-Neon laser (633nm) and 660/20 bandpass filter.

  • Alexa Fluor® 647 Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab237478)
    Alexa Fluor® 647 Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab237478)

Protocols

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

References (0)

Publishing research using ab237478? Please let us know so that we can cite the reference in this datasheet.

ab237478 has not yet been referenced specifically in any publications.

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