Recombinant
RabMAb

Anti-Histone H4 (mono methyl K20) antibody [EPR16999] - ChIP Grade (ab177188)

Overview

  • Product name
    Anti-Histone H4 (mono methyl K20) antibody [EPR16999] - ChIP Grade
    See all Histone H4 primary antibodies
  • Description
    Rabbit monoclonal [EPR16999] to Histone H4 (mono methyl K20) - ChIP Grade
  • Host species
    Rabbit
  • Tested applications
    Suitable for: PepArr, ICC/IF, ChIP, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Histone H4 aa 1-100 (mono methyl K20). The exact sequence is proprietary.
    Database link: P62805

  • Positive control
    • ICC/IF: HeLa cells. IHC-P: Human and mouse colon tissues; Rat kidney tissue. ChIP: Chromatin from HeLa cells. WB: HeLa whole cell lysates; NIH3T3 nuclear extracts lysates.
  • 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
    EPR16999
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab177188 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.
ICC/IF 1/100.
ChIP Use 2 µg for 25 µg of chromatin.
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/5000. Detects a band of approximately 11 kDa (predicted molecular weight: 11 kDa).

Target

Images

  • Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25μg of chromatin, 2μg of ab177188 (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 approach). Primers are located in the first kb of the transcribed region.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling Histone H4 (mono methyl K20) with ab177188 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Nuclear staining on HeLa cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:
    1. ab177188 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

     

  • All lanes : Anti-Histone H4 (mono methyl K20) antibody [EPR16999] - ChIP Grade (ab177188) at 1/5000 dilution

    Lane 1 : Hela whole cell lysates
    Lane 2 : NIH/3T3 whole cell lysates

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 11 kDa
    Observed band size: 11 kDa



    Blocking buffer/Diluting buffer: 5% NFDM/TBST

  • ab177188 was 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.

  • Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Histone H4 (mono methyl K20) with ab177188 at 1/100 dilution, followed by prediluted Goat Anti-Rabbit IgG H&L (HRP). Nucleus staining on glandular epithelium of colon tissue is observed. Counter stained with Hematoxylin.

    Negative control: PBS instead of primary antibody, secondary antibody is prediluted Goat Anti-Rabbit IgG H&L (HRP).

  • Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Histone H4 (mono methyl K20) with ab177188 at 1/100 dilution, followed by prediluted Goat Anti-Rabbit IgG H&L (HRP). Nucleus staining on glandular epithelium of mouse colon tissue is observed. Counter stained with Hematoxylin.

    Negative control: PBS instead of primary antibody, secondary antibody is prediluted Goat Anti-Rabbit IgG H&L (HRP).

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Histone H4 (mono methyl K20) with ab177188 at 1/100 dilution, followed by prediluted Goat Anti-Rabbit IgG H&L (HRP). Nucleus staining on rat kidney tissue is observed. Counter stained with Hematoxylin.

    Negative control: PBS instead of primary antibody, secondary antibody is prediluted Goat Anti-Rabbit IgG H&L (HRP).

References

ab177188 has not yet been referenced specifically in any publications.

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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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