Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (ab17036)

Overview

  • Product name
    Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade
    See all Histone H4 primary antibodies
  • Description
    Mouse monoclonal [mAbcam 17036] to Histone H4 - ChIP Grade
  • Tested applications
    Suitable for: IHC-P, ChIP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
  • Immunogen

    Synthetic peptide corresponding to Human Histone H4 aa 50 to the C-terminus (C terminal) conjugated to Keyhole Limpet Haemocyanin (KLH).
    Database link: P62805
    (Peptide available as ab13843)

  • Positive control
    • This antibody gave a positive signal in the following: Calf thymus histone preparation; HeLa histone preparation; Histone H4 recombinant protein; NIH3T3 Whole Cell Lysate; MEF1 Whole Cell Lysate; PC12 Whole Cell Lysate.
  • General notes

    This antibody clone is manufactured by Abcam.

    To detect histone H4 by ICC/IF, we recommend using ab177840 or ab9052 since these antibodies perform better in this application.

Properties

Applications

Our Abpromise guarantee covers the use of ab17036 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.
ChIP Use 5 µg for 25 µg of chromatin.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 14 kDa (predicted molecular weight: 11 kDa).Can be blocked with Human Histone H4 peptide (ab13843).

We recommend using 3% milk as the blocking agent for Western blot.

Target

Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade images

  • All lanes : Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (ab17036) at 2 µg/ml

    Lane 1 : Histone H1 Recombinant Protein
    Lane 2 : Histone H2A Recombinant Protein
    Lane 3 : Histone H2B Recombinant Protein
    Lane 4 : Histone H3.1 Recombinant Protein
    Lane 5 : Histone H4 Recombinant Protein

    Lysates/proteins at 0.1 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution

    Performed under reducing conditions.

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


    Exposure time : 30 seconds
  • All lanes : Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (ab17036) at 5 µg/ml

    Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg
    Lane 2 : HeLa Histone Preparation Nuclear Lysate at 2.5 µg

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

    Performed under reducing conditions.

    Predicted band size : 11 kDa
    Observed band size : 14 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute
  • All lanes : Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (ab17036) at 1 µg/ml

    Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 11 kDa
    Observed band size : 14 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute
  • Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab17036 (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 for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.
  • IHC image of Histone H4 staining in formalin fixed, paraffin embedded human Hodgkin's lymphoma tissue section*, performed on a Leica Bond™ 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 ab17036, 1 µg/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.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • IHC image of ab17036 staining in Human Hodgekin's Lymphoma 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 ab17036, 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.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  • All lanes : Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (ab17036) at 5 µg/ml

    Lane 1 : Histone H1 Recombinant Protein
    Lane 2 : Histone H2A Recombinant Protein
    Lane 3 : Histone H2B Recombinant Protein
    Lane 4 : Histone H3.1 Recombinant Protein
    Lane 5 : Histone H4 Recombinant Protein

    Lysates/proteins at 0.1 µg per lane.

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

    Performed under reducing conditions.

    Predicted band size : 11 kDa
    Observed band size : 14 kDa (why is the actual band size different from the predicted?)


    Exposure time : 30 seconds

References for Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (ab17036)

This product has been referenced in:
  • Walsh RM  et al. Phf8 loss confers resistance to depression-like and anxiety-like behaviors in mice. Nat Commun 8:15142 (2017). WB ; Mouse . Read more (PubMed: 28485378) »
  • Dikopoltsev E  et al. FBXO22 protein is required for optimal synthesis of the N-methyl-D-aspartate (NMDA) receptor coagonist D-serine. J Biol Chem 289:33904-15 (2014). Read more (PubMed: 25336657) »

See all 2 Publications for this product

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

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