Overview

  • Product name
    Anti-Histone H4 (di methyl K79) antibody
    See all Histone H4 primary antibodies
  • Description
    Rabbit polyclonal to Histone H4 (di methyl K79)
  • Host species
    Rabbit
  • Specificity
    Specific to Histone H4 K79 Di-methyl. In blocking experiments, is not blocked by H3 K79 peptides.
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, ICCmore details
  • Species reactivity
    Reacts with: Cow, Human
    Does not react with: Saccharomyces cerevisiae
  • Immunogen

    Synthetic peptide within Human Histone H4 aa 50 to the C-terminus (di methyl K79) conjugated to Keyhole Limpet Haemocyanin (KLH). The exact sequence is proprietary.
    (Peptide available as ab4560)

  • Positive control
    • Calf Thymus Histone Preparation; Hela whole cell extract

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab2885 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 1/500. Detects a band of approximately 14 kDa.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/200.
ICC 1/200.

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 H4 family.
  • Post-translational
    modifications
    Acetylation at Lys-6 (H4K5ac), Lys-9 (H4K8ac), Lys-13 (H4K12ac) and Lys-17 (H4K16ac) occurs in coding regions of the genome but not in heterochromatin.
    Citrullination at Arg-4 (H4R3ci) by PADI4 impairs methylation.
    Monomethylation and asymmetric dimethylation at Arg-4 (H4R3me1 and H4R3me2a, respectively) by PRMT1 favors acetylation at Lys-9 (H4K8ac) and Lys-13 (H4K12ac). Demethylation is performed by JMJD6. Symmetric dimethylation on Arg-4 (H4R3me2s) by the PRDM1/PRMT5 complex may play a crucial role in the germ-cell lineage.
    Monomethylated, dimethylated or trimethylated at Lys-21 (H4K20me1, H4K20me2, H4K20me3). Monomethylation is performed by SET8. Trimethylation is performed by SUV420H1 and SUV420H2 and induces gene silencing.
    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. Monoubiquitinated at Lys-92 of histone H4 (H4K91ub1) in response to DNA damage. The exact role of H4K91ub1 in DNA damage response is still unclear but it may function as a licensing signal for additional histone H4 post-translational modifications such as H4 Lys-21 methylation (H4K20me).
    Sumoylated, which is associated with transcriptional repression.
  • Cellular localization
    Nucleus. Chromosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • dJ160A22.1 antibody
    • dJ160A22.2 antibody
    • dJ221C16.1 antibody
    • dJ221C16.9 antibody
    • FO108 antibody
    • H4 antibody
    • H4.k antibody
    • H4/a antibody
    • H4/b antibody
    • H4/c antibody
    • H4/d antibody
    • H4/e antibody
    • H4/g antibody
    • H4/h antibody
    • H4/I antibody
    • H4/j antibody
    • H4/k antibody
    • H4/m antibody
    • H4/n antibody
    • H4/p antibody
    • H4_HUMAN antibody
    • H4F2 antibody
    • H4F2iii antibody
    • H4F2iv antibody
    • H4FA antibody
    • H4FB antibody
    • H4FC antibody
    • H4FD antibody
    • H4FE antibody
    • H4FG antibody
    • H4FH antibody
    • H4FI antibody
    • H4FJ antibody
    • H4FK antibody
    • H4FM antibody
    • H4FN antibody
    • H4M antibody
    • HIST1H4A antibody
    • HIST1H4B antibody
    • HIST1H4C antibody
    • HIST1H4D antibody
    • HIST1H4E antibody
    • HIST1H4F antibody
    • HIST1H4H antibody
    • HIST1H4I antibody
    • HIST1H4J antibody
    • HIST1H4K antibody
    • HIST1H4L antibody
    • HIST2H4 antibody
    • HIST2H4A antibody
    • Hist4h4 antibody
    • Histone 1 H4a antibody
    • Histone 1 H4b antibody
    • Histone 1 H4c antibody
    • Histone 1 H4d antibody
    • Histone 1 H4e antibody
    • Histone 1 H4f antibody
    • Histone 1 H4h antibody
    • Histone 1 H4i antibody
    • Histone 1 H4j antibody
    • Histone 1 H4k antibody
    • Histone 1 H4l antibody
    • Histone 2 H4a antibody
    • histone 4 H4 antibody
    • Histone H4 antibody
    • MGC24116 antibody
    see all

Images

  • All lanes : Anti-Histone H4 (di methyl K79) antibody (ab2885) at 1/500 dilution

    Lane 1 : Calf Thymus histone prep
    Lane 2 : Calf Thymus histone prep with Human Histone H3 (mono methyl K79) peptide (ab4555) at 0.1 µg
    Lane 3 : Calf Thymus histone prep with Human Histone H3 (di methyl K79) peptide (ab4556) at 0.1 µg
    Lane 4 : Calf Thymus histone prep with Human Histone H3 (tri methyl K79) peptide (ab4557) at 0.1 µg
    Lane 5 : Calf Thymus histone prep with Human Histone H4 (di methyl K79) peptide (ab4560) at 0.1 µg
    Lane 6 : Calf Thymus histone prep with Human Histone H3 (di methyl K27) peptide (ab1781) at 0.1 µg

    Lysates/proteins at 1 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/2000 dilution


    Rabbit polyclonal to Histone H4 Di Methyl K79 at 1/500 on Calf Thymus histone prep (1 ug per lane).

    Lane 1: ab2885 1/500
    Lane 2: ab2885 1/500 H3 Mono Me K79 peptide (ab4555), 0.1 ug
    Lane 3: ab2885 1/500 H3 Di Me K79 peptide (ab4556), 0.1 ug
    Lane 4: ab2885 1/500 H3 Tri Me K79 peptide  (ab4557), 0.1 ug
    Lane 5: ab2885 1/500 H4 Di Me K79 peptide  (ab4560), 0.1 ug
    Lane 6: ab2885 1/500 H3 Di Me K27 peptide  (ab1781), 0.1 ug

    Secondary antibody: Goat anti-rabbit IgG (HRP) ab 1/2000 (ab6721)

  • SKN cells were stained with ab2885 (green) at a dilution of 1/200. The cells were fixed in paraformaldehyde for 10 minutes prior to incubation with ab2885. The DNA was stained with DAPI (blue). 100x magnification.

  • ab2885 at 1/2000 used in Western blot analysis of MCF7 cells (human breast cancer cells).

    Lane 1: MCF7 cell lysate  control

    Lane 2: MCF7 cell lysate (cells  treated with 50µm Troglitazone for 48 hours)

    Lane 3: MCF7 cells lysate (cells treated with 1µm Trichostatin for 48 hours)

    Troglitazone and trichostatin increase dimethyl H4 (K79) and act as anti-proliferatives in human breast cancer. The blot was stripped and reprobed with an alpha tubilin antibody to show loading equivalence.

    See Abreview

  • IHC image of Histone H4 (di methyl K79) staining in human breast carcinoma FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab2885, 5µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

References

This product has been referenced in:
  • Zhang K  et al. Comparative analysis of histone H3 and H4 post-translational modifications of esophageal squamous cell carcinoma with different invasive capabilities. J Proteomics 112C:180-189 (2014). Read more (PubMed: 25234497) »
See 1 Publication for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Saccharomyces cerevisiae Purified protein (Acid extracted histones)
Loading amount
3 µg
Specification
Acid extracted histones
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 12 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (human MCF 7 breast cancer cells)
Loading amount
10 µg
Specification
human MCF 7 breast cancer cells
Treatment
50uM TRG and 1uM Trichostatin A for 48 hours
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Dr. Gerald Davies

Verified customer

Submitted Oct 05 2006

Answer

Thank you for your query regarding ab2885. The western blot image on the datasheet was performed on purified histone preps from Calf thymus with 1 ug loading per lane (a large excess of protein). The top band relates to H3 with H2A and H2B between these and the lowest band which corresponds to H4. The MW markers on this image do appear to indicate that the H4 runs at a higher MW than would be expected but I consider this to be a fault in the resolution of these markers at the low MW range. As we did use purified histones there can be little else but H4 running at the bottom band and as the image shows this is specifically blocked by Di Me K79 H4 blocking peptide. The detection of the H3 band is probably due to the high levels of protein on the membrane. I am therefore confident that the antibody detects H4 and we shall investigate this blot with more accurate MW markers to improve the image in future. Thank you for alerting us to this and I hope that we have addressed your concerns satisfactorily.

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up