Anti-Histone H4 (di methyl K20, tri methyl K20) antibody [6F8-D9] (ab78517)

Overview

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
    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity
    IgG fraction
  • Clonality
    Monoclonal
  • Clone number
    6F8-D9
  • Myeloma
    x63-Ag8.653
  • Isotype
    IgG1
  • Light chain type
    kappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab78517 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
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use at an assay dependent concentration. Predicted molecular weight: 11 kDa.
ELISA Use at an assay dependent concentration.
Dot blot Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Target

Anti-Histone H4 (di methyl K20, tri methyl K20) antibody [6F8-D9] images

  • ICC/IF image of ab78517 stained mouse oocyte. Oocytes were fixed in formaldehyde and then incubated in 3% serum with 0.25% Triton for 30 minutes at 37°C. Cells were incubated with the antibody (ab78517) at 1/200 for 2 hours at 37°C. The secondary antibody used was a polyclonal Goat-anti-mouse conjugated to Alexa Fluor® 488 (1/250).

    See Abreview

  • All lanes : Anti-Histone H4 (di methyl K20, tri methyl K20) antibody [6F8-D9] (ab78517) at 1 µg/ml

    Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate
    Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H4 peptide (ab14963) at 0.5 µg/ml
    Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K4) peptide (ab1340) at 0.5 µg/ml
    Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K4) peptide (ab7768) at 0.5 µg/ml
    Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K4) peptide (ab1342) at 0.5 µg/ml
    Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H4 (mono methyl K20) peptide (ab17043) at 0.5 µg/ml
    Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H4 peptide (17-24) - di methyl K20 at 0.5 µg/ml
    Lane 8 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H4 (tri methyl K20) peptide (ab17567) at 0.5 µg/ml
    Lane 9 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K9) peptide (ab1771) at 0.5 µg/ml
    Lane 10 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µg/ml
    Lane 11 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K9) peptide (ab1773) at 0.5 µg/ml

    Lysates/proteins at 0.5 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 11 kDa
    Observed band size : 13 kDa (why is the actual band size different from the predicted?)


    Exposure time : 90 secondsab78517 is specific for Histone H4 (tri-methyl K20). This is illustrated in lane 8 where the activity of ab78517 is specifically blocked by the addition of the immunizing peptide (ab17567).
  • ICC/IF image of ab78517 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab78517, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) Hek293, HepG2 and MCF7 cells at 5µg/ml, and in 4% PFA fixed (10 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml.
  • IHC image of Histone H4 (tri methyl K20) staining in normal human skin formalin fixed paraffin embedded tissue section, performed on a Leica 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 ab78517, 1µg/ml, for 15 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.
  • Overlay histogram showing HeLa cells stained with ab78517 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab78517, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed.

References for Anti-Histone H4 (di methyl K20, tri methyl K20) antibody [6F8-D9] (ab78517)

This product has been referenced in:
  • Tikhmyanova N  et al. Small molecule perturbation of the CAND1-Cullin1-ubiquitin cycle stabilizes p53 and triggers Epstein-Barr virus reactivation. PLoS Pathog 13:e1006517 (2017). Read more (PubMed: 28715492) »
  • Upadhyay M  et al. Transposon Dysregulation Modulates dWnt4 Signaling to Control Germline Stem Cell Differentiation in Drosophila. PLoS Genet 12:e1005918 (2016). Read more (PubMed: 27019121) »

See all 8 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Oocyte)
Specification
Oocyte
Fixative
Formaldehyde
Permeabilization
Yes - 0.25% Triton
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 37°C
Username

Dr. Maki Asami

Verified customer

Submitted May 09 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Caenorhabditis elegans Cell (Intestinal Nuclei)
Specification
Intestinal Nuclei
Fixative
Paraformaldehyde
Permeabilization
Yes - Sperm Salts Buffer, Triton X-100
Username

Abcam user community

Verified customer

Submitted Jun 06 2011

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

Sign up