Anti-Histone H4 antibody - ChIP Grade (ab7311)

Overview

  • Product nameAnti-Histone H4 antibody - ChIP Grade
    See all Histone H4 primary antibodies
  • Description
    Rabbit polyclonal to Histone H4 - ChIP Grade
  • SpecificityHistone H4
  • Tested applicationsSuitable for: IHC-P, ICC/IF, ChIP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Cow, Human, Saccharomyces cerevisiae
    Predicted to work with: all Mammals
  • Immunogen

    Synthetic peptide corresponding to Human Histone H4 aa 1-100 conjugated to Keyhole Limpet Haemocyanin (KLH).
    Database link: P62805
    (Peptide available as ab21044)

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

Properties

Applications

Our Abpromise guarantee covers the use of ab7311 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.
ICC/IF 1/200.
ChIP Use 2 µg for 25 µg of chromatin.
WB 1/500 - 1/1000. Detects a band of approximately 14 kDa (predicted molecular weight: 11 kDa).

Target

Anti-Histone H4 antibody - ChIP Grade images

  • All lanes : Anti-Histone H4 antibody - ChIP Grade (ab7311) at 1/500 dilution

    Lane 1 : Calf thymus histone extract
    Lane 2 : Calf thymus histone extract with Human Histone H4 peptide (ab21044) at 1 µg/ml
    Lane 3 : Calf thymus histone extract with Histone H4 peptide - acetyl K16 (ab22376) at 1 µg/ml
    Lane 4 : Calf thymus histone extract with Human Histone H4 (acetyl K20) peptide (ab20632) at 1 µg/ml

    Lysates/proteins at 0.5 µg per lane.

    Secondary
    Alexa-Fluor Goat anti-rabbit IgG at 1/10000 dilution

    Predicted band size : 11 kDa
    Observed band size : 14 kDa (why is the actual band size different from the predicted?)
  • 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 ab7311 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.

  • IHC image of Histone H4 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 ab7311, 1µg/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.
  • SKN-SH cells were fixed in 4% paraformaldehyde for 10 mins, permeabilized in PBS-0.5% Triton X-100 for 5 mins and incubated for 30 minutes with ab7311 (1/200 dilution). The slides were rinsed once in PBS-Triton (0.1%), twice in PBS then incubated with the secondary antibody for 30 mins. The DNA is stained with DAPI (blue). Clear nuclear staining with ab7311 can be seen (green).

References for Anti-Histone H4 antibody - ChIP Grade (ab7311)

This product has been referenced in:
  • Westhorpe FG  et al. A cell-free CENP-A assembly system defines the chromatin requirements for centromere maintenance. J Cell Biol 209:789-801 (2015). Read more (PubMed: 26076692) »
  • Ghule PN  et al. Fidelity of histone gene regulation is obligatory for genome replication and stability. Mol Cell Biol 34:2650-9 (2014). Read more (PubMed: 24797072) »

See all 25 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (multiple myeloma cell)
Loading amount 50 µg
Specification multiple myeloma cell
Gel Running Conditions Reduced Denaturing (10-20% tris glycine gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Nov 21 2012

Thanks in advance for filling out the questionnaire below for each antibody. The answers will help me to assist you most efficiently.

1) Abcam product code ab



2) Abcam order reference number or product batch number
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Thanks for your email.

This item is currently back ordered 1-2 weeks from date the order is placed. You should expect to receive this vial of ab64171 in approximately 1 week.

I hope this is helpful. Please contact us again if you ha...

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Fantastic! Best of luck in your experiments!

Thank you for your enquiry.

We have a number of different antibodies against specific acetylated residues on histone H4. However an antibody does not exist that will detect every acetylated residue on histone H4.

You may consider ju...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Tissue lysate - whole (Bone tissue)
Loading amount 25 µg
Specification Bone tissue
Gel Running Conditions Reduced Denaturing (15% Gel)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Dr. Rodrigo Alves

Verified customer

Submitted Apr 17 2012

DISCOUNT CODE: 100%ABR-T7A9Q
Expiration date: 12th August 2012

I am very pleased to hear you would like to accept our offer and test ab7311 insheep tissue. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To r...

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I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so ...

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Thanks for your reply.

No, there is no need to return the unused portion of the antibody to Abcam.

I am able to set up the refund myself. You do not need to fill out any forms. I just need to locate the order in our system to proces...

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Thank you for your reply.

I would be happy to arrange a refund for your recent purchase of ab117635. If you could, please forward me the purchase order number or Abcam order reference number associated with this purchase so I can initiate the...

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