Overview

  • Product name
  • Description
    Chicken polyclonal to Histone H4
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken, Cow, Pig, Drosophila melanogaster, Zebrafish
  • 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
    • WB: Recombinant Histone H4 protein and NIH3T3, MEF1, PC12 and HeLa cell lysates and HeLa Histone preparation. ICC/IF: methanol fixed HeLa cells. IHC-P: Human breast fibroadenoma tissue.

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
    Constituents: PBS, 3% BSA

    This product may contain up to 3% BSA depending on the batch. For specific batch formulations please contact us.
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgY
  • Research areas

Associated products

Applications

Our Abpromise guarantee covers the use of ab134212 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 14 kDa (predicted molecular weight: 11 kDa).
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

Images

  • All lanes : Anti-Histone H4 antibody (ab134212) at 1 µg/ml

    Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg
    Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µg
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
    Lane 5 : HeLa Histone Preparation Nuclear Lysate at 2.5 µg
    Lane 6 : Histone H4 Recombinant Protein at 0.1 µg
    Lane 7 : Histone H3.1 Recombinant Protein (negative control) at 0.1 µg

    Secondary
    Goat polyclonal Secondary Antibody to Chicken IgY - H&L (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 11 kDa


    Exposure time : 3 minutes
  • All lanes : Anti-Histone H4 antibody (ab134212) at 1 µg/ml

    Lane 1 : Histone H4 Recombinant Protein
    Lane 2 : Histone H2A Recombinant Protein

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal Secondary Antibody to Chicken IgY - H&L (HRP) at 1/3000 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 : 8 minutes
  • ICC/IF image of ab134212 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 ab134212 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- chicken IgY (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.

  • IHC image of Histone H4 staining in human breast fibroadenoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. 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 ab134212, 5µg/ml, for 15 mins at room temperature. A goat anti-chicken biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC 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.

References

This product has been referenced in:
  • Ganatra HA  et al. Zinc supplementation leads to immune modulation and improved survival in a juvenile model of murine sepsis. Innate Immun 23:67-76 (2017). Read more (PubMed: 27821649) »

See 1 Publication 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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