Recombinant
RabMAb

Recombinant Anti-Histone H3 (citrulline R26) antibody [EPR20606] (ab212082)

Overview

  • Product name

    Anti-Histone H3 (citrulline R26) antibody [EPR20606]
    See all Histone H3 primary antibodies
  • Description

    Rabbit monoclonal [EPR20606] to Histone H3 (citrulline R26)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Dot blot, ELISAmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    This product was produced with the following immunogens:
    Synthetic peptide within Human Histone H3 aa 1-100 (citrulline R26). The exact sequence is proprietary.
    Database link: P68431

    Synthetic peptide within Human Histone H3 aa 1-100 (citrulline R26). The exact sequence is proprietary.
    Database link: P68431

  • Positive control

    • WB: HEK-293T transfected with GFP-tagged PADI4 (WT) whole cell lysate treated with CaCl2 and Ionomycin; NIH/3T3 transfected with GFP-tagged PADI4 (WT) whole cell lysate treated with CaCl2.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 0.05% BSA, 40% Glycerol, PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20606
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab212082 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
WB 1/1000. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).
Dot blot 1/1000.
ELISA Use a concentration of 0 - 1 µg/ml.

Target

  • 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.
  • Information by UniProt
  • Database links

  • 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

Images

  • All lanes : Anti-Histone H3 (citrulline R26) antibody [EPR20606] (ab212082) at 1/1000 dilution

    Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with an empty vector (control) then treated with 10 mM CaCl2 and 10 µM Ionomycin for 2 hours, whole cell lysate
    Lane 2 : HEK-293T transfected with a GFP-tagged PADI4 expression vector then treated with 10 mM CaCl2 and 10 µM Ionomycin for 2 hours, whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

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


    Exposure time: 5 seconds


    Histone H3R26 is citrullinated by PADI4, CaCl2 is used as a co-factor according to the literature (PMID: 16567635). Ionomycin is used to improve the modification by PADI4 according to the literature (PMID: 26360112).

    Blocking/Dilution buffer: 5% BSA/TBST.

  • Dot blot analysis of Histone H3 (citrulline R26) labeled with ab212082 at 1/1000 dilution.

    Lane 1: Histone H3 (citrulline R26) peptide (aa25-36)

    Lane 2: Histone H3 (Citrulline R17) peptide (aa1-21)

    Lane 3: Histone H3 (Citrulline R8) peptide (aa1-21)

    Lane 4: Histone H3 (Citrulline R2) peptide (aa1-21)

    Lane 5: Histone H3 peptide (aa1-21)

    Lane 6: Histone H3 peptide (aa19-36)

    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 30 seconds.

  • All lanes : Anti-Histone H3 (citrulline R26) antibody [EPR20606] (ab212082) at 1/1000 dilution

    Lane 1 : NIH/3T3 (mouse embryonic fibroblast cell line) transfected with a GFP-tagged empty vector (control) then treated with 10 mM CaCl2 for 2 hours, whole cell lysate
    Lane 2 : NIH/3T3 transfected with a GFP-tagged PADI4 expression vector treated with 10 mM CaCl2 for 2 hours, whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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


    Exposure time: 3 minutes


    Histone H3R26 is citrullinated by PADI4, CaCl2 is used as a co-factor according to the literature (PMID: 16567635). Ionomycin is used to improve the modification by PADI4 according to the literature (PMID: 26360112).

    Blocking/Dilution buffer: 5% BSA/TBST.

  • ELISA using ab212082 labeling Histone H3 (Citrulline R26), Histone H3R26 unmodified, Histone H3 (Citrulline R17), Histone H3 (Citrulline R8), Histone H3 (Citrulline R2) and Histone H3R2/8/17 unmodified at 1 µg/ml. ab212082 was used at a range between 0 -1 µg/ml.

    Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) was used as secondary antibody at 1/2500 dilution.

    ab212082 cross binds H3R8Cit at 14%.

References

ab212082 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
ChIP
Sample
Human Cell lysate - nuclear (MCF7 breast cancer cell line)
Negative control
Rabbit IgG
Specification
MCF7 breast cancer cell line
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 0.75% PFA
Positive control
Estrogen receptor antibody for ChIP, H3Cit2/8/17 antibody for ChIP Our previous publication which used other H3Cit26 antibody that has now been discontinued. (Proceedings of the National Academy of Sciences Aug 2012, 109 (33) 13331-13336 DOI: 10.1073/pnas.1203280109)

Abcam user community

Verified customer

Submitted Aug 02 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (MCF7 breast cancer cells)
Permeabilization
Yes - Triton X-100
Specification
MCF7 breast cancer cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 09 2018

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