Recombinant
RabMAb

Anti-Histone H2B (acetyl K20) antibody [EPR859] - ChIP Grade (ab177430)

Overview

  • Product name
    Anti-Histone H2B (acetyl K20) antibody [EPR859] - ChIP Grade
    See all Histone H2B primary antibodies
  • Description
    Rabbit monoclonal [EPR859] to Histone H2B (acetyl K20) - ChIP Grade
  • Host species
    Rabbit
  • Tested applications
    Suitable for: PepArr, IHC-P, WB, ICC/IF, Flow Cyt, ChIPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Histone H2B aa 1-100 (acetyl K20). The exact sequence is proprietary.
    Database link: Q16778

  • Positive control
    • WB: HeLa and NIH/3T3 whole cell lysate treated with 500 ng/ml Trichostatin A for 4 hours. IHC: Human and rat colon tissue, Mouse kidney tissue. ICC/IF/FC: HeLa cells
  • 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

Applications

Our Abpromise guarantee covers the use of ab177430 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
PepArr Use at an assay dependent concentration.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa).
ICC/IF 1/2000.
Flow Cyt 1/300.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

ChIP Use 2 µg for 25 µg of chromatin.

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 H2B family.
  • Post-translational
    modifications
    Monoubiquitination of Lys-121 by the RNF20/40 complex gives a specific tag for epigenetic transcriptional activation and is also prerequisite for histone H3 'Lys-4' and 'Lys-79' methylation. It also functions cooperatively with the FACT dimer to stimulate elongation by RNA polymerase II.
    Phosphorylated on Ser-15 by STK4/MST1 during apoptosis; which facilitates apoptotic chromatin condensation. Also phosphorylated on Ser-15 in response to DNA double strand breaks (DSBs), and in correlation with somatic hypermutation and immunoglobulin class-switch recombination.
  • Cellular localization
    Nucleus. Chromosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • H2B 1A antibody
    • H2B antibody
    • H2B histone family antibody
    • H2B2f antibody
    • H2B2F_HUMAN antibody
    • H2Ba antibody
    • H2Bf antibody
    • HIST2H2BF antibody
    • histone H2B antibody
    • histone H2B type 1 antibody
    • Histone H2B type 2-F antibody
    • MGC131639 antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Histone H2B (acetyl K20) with ab177430 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on Human colon is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary antibody.

  • All lanes : Anti-Histone H2B (acetyl K20) antibody [EPR859] - ChIP Grade (ab177430) at 1/10000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) treated with 500 ng/ml Trichostatin A for 4 hours whole cell lysates
    Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) untreated whole cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated

    Predicted band size: 14 kDa
    Observed band size: 14 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells, treated with Trichostatin A (500 ng/ml) for 4 hours or untreated, labeling Histone H2B (acetyl K20)  with ab177430 at 1/2000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on HeLa cell line is observed. Acetylation level increased after treatment with Trichostatin A (500 ng/ml) for 4 hours. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows;
    1. ab177430 at 1/2000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • All batches of ab177430 are tested in Peptide Array against 501 different modified and unmodified histone peptides; each peptide is printed on the array at six concentrations (each in triplicate).
     
    Circle area represents affinity between the antibody and a peptide: all antigen-containing peptides are displayed as red circles, all other peptides as blue circles. The affinity is calculated as area under curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity.
     
    The complete dataset, including full list of all peptides and information on the position of each peptide in the diagram, can be downloaded here.

  • All lanes : Anti-Histone H2B (acetyl K20) antibody [EPR859] - ChIP Grade (ab177430) at 1/1000 dilution

    Lane 1 : NIH/3T3 (Mouse embyro fibroblast cells) treated with 500 ng/ml Trichostatin A for 4 hours whole cell lysates 10ug
    Lane 2 : NIH/3T3 (Mouse embyro fibroblast cells) untreated whole cell lysates 10ug

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 14 kDa
    Observed band size: 14 kDa


    Exposure time: 10 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Histone H2B (acetyl K20) with ab177430 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on mouse kidney is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary antibody.

  • Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Histone H2B (acetyl K20) with ab177430 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on Rat colon is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary antibody.

  • Flow cytometric analysis of 2% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells treated with 500ng/ml Trichostatin A for 4 hours, labeling Histone H2B (acetyl K20) with ab177430 at 1/300 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

  • Chromatin was prepared from HeLa (Human epithelial cells from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab177430 (blue), and 20µl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach). Primers and probes are located in the first kb of the transcribed region.

References

This product has been referenced in:
  • Sainz de la Maza D  et al. Metabolic Reprogramming, Autophagy, and Reactive Oxygen Species Are Necessary for Primordial Germ Cell Reprogramming into Pluripotency. Oxid Med Cell Longev 2017:4745252 (2017). ICC/IF ; Mouse . Read more (PubMed: 28757909) »
  • Rothbart SB  et al. An Interactive Database for the Assessment of Histone Antibody Specificity. Mol Cell 59:502-11 (2015). Read more (PubMed: 26212453) »

See all 2 Publications for this product

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

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