Recombinant
RabMAb

Recombinant Anti-Histone H3 (ubiquityl K23) antibody [EPR18876] (ab194243)

Overview

  • Product name

    Anti-Histone H3 (ubiquityl K23) antibody [EPR18876]
    See all Histone H3 primary antibodies
  • Description

    Rabbit monoclonal [EPR18876] to Histone H3 (ubiquityl K23)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Dot blot, IP, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Histone H3 aa 1-100 (ubiquityl K23). The exact sequence is proprietary.
    Database link: P68431

  • Positive control

    • WB: Lysate of HeLa cells treated with 2mM thymidine for 18 hrs, followed by addition of fresh culture medium and incubation for 9 hrs, then treated with 2mM thymidine for 15 hrs, followed by addition of fresh culture medium and incubation for 4 hrs; Jurkat, HEK-293 lysates. IP: Lysate of HeLa cells treated with 2mM thymidine for 18 hrs, followed by addition of fresh culture medium and incubation for 9 hrs, then 2mM thymidine for 15 hrs, followed by addition of fresh culture medium and incubation for 4 hrs.
  • 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: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR18876
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab194243 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
Dot blot Use at an assay dependent concentration.
IP 1/30.
WB 1/1000. Detects a band of approximately 28,35 kDa (predicted molecular weight: 15 kDa).

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 (ubiquityl K23) antibody [EPR18876] (ab194243) at 1/20000 dilution

    Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : Whole cell lysates from HeLa treated with 2mM thymidine for 18 hours, followed by addition of fresh culture medium and incubation for 9 hours. Cell were then treated with 2mM thymidine for 15 hours, followed by addition of fresh culture medium and incubation for 4 hours

    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: 28 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression level of H3K23ubi is increased during S phase. PMID 24013172.

    The MW is consistent with literature: PMID 24013172.

  • Histone H3 (ubiquityl K23) was immunoprecipitated using ab194243 at 1/30 dilution from 0.35 mg of whole cell lysate from HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 2mM thymidine for 18 hours, followed by addition of fresh culture medium and incubation for 9 hours, then treated with 2mM thymidine for 15 hours, followed by addition of fresh culture medium and incubation for 4 hours.

    Western blot was performed from the immunoprecipitate using ab194243 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: 10 µg of whole cell lysate from HeLa treated with 2mM thymidine for 18 hours, followed by addition of fresh culture medium and incubation for 9 hours, cells were then treated with 2mM thymidine for 15 hours, followed by addition of fresh culture medium and incubation for 4 hours (Input).

    Lane 2: ab194243 IP in whole cell lysate from HeLa treated with 2mM thymidine for 18 hours, followed by addition of fresh culture medium and incubation for 9 hours, cells were then treated with 2mM thymidine for 15 hours, followed by addition of fresh culture medium and incubation for 4 hours.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab194243 in whole cell lysate from HeLa treated with 2mM thymidine for 18 hours, followed by addition of fresh culture medium and incubation for 9 hours, cells were then treated with 2mM thymidine for 15 hours, followed by addition of fresh culture medium and incubation for 4 hours.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 seconds.

    The band at around 35kDa can be explained as di-ubiquitinated K23 (PMID24013172).

  • All lanes : Anti-Histone H3 (ubiquityl K23) antibody [EPR18876] (ab194243) at 1/20000 dilution

    Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
    Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate

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

    Predicted band size: 15 kDa
    Observed band size: 28 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The MW is consistent with literature: PMID 24013172.

  • Dot blot analysis of Histone H3 (ubiquityl K23) labeled with ab194243 at 1/1000 dilution.

    Lane 1: Histone H3 (ubiquitinated K23) peptide (aa20-29).

    Lane 2: Histone H3 non- ubiquitinated peptide (aa22-31).

    Lane 3: Histone H3 full length recombinant protein.

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

    Blocking and diluting buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

References

ab194243 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab194243.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up