Overview

  • Product name
  • Description
    Rabbit polyclonal to Histone H4
  • Specificity
    ab52178 detects endogenous levels of total Histone H4 protein
  • Tested applications
    Suitable for: ELISA, ICC/IF, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide corresponding to Human Histone H4 (N terminal). Immunogen is in the range of aa 15-64.
    Database link: P62805

  • Positive control
    • WB: Extracts from RAW264.7 cells, treated with TSA (400nM, 24hours). IHC-P: Human normal skin tissue. ICC/IF: HeLa cells.

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. Stable for 12 months at -20°C.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 50% Glycerol, PBS (without Mg2+ and Ca2+), 150mM Sodium chloride, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    ab52178 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab52178 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
ELISA 1/20000.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/500 - 1/1000. Predicted molecular weight: 11 kDa.

Target

Images

  • ab52178 stained HeLa cells. The cells were serum starved and then fixed using 100% methanol for 5 minutes at -20°C then permeabilised using 0.1% PBS-Triton. The cells were then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to further permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52178 at 1µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Ab195889 Mouse monoclonal [DM1A] to alpha Tubulin - Microtubule Marker (Alexa Fluor® 594) was used to label microtubules (pseudo-colored red) at a 1/250 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • ab52178 stained HeLa cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52178 at 1µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • IHC image of Histone H4 staining in Human normal skin formalin fixed paraffin embedded 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 ab52178, 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 (ab52178) at 1/500 dilution

    Lane 1 : RAW264.7 cell extract, treated with TSA (400nM, 24hours)
    Lane 2 : RAW264.7 cell extract, treated with TSA (400nM, 24hours) with immunising peptide


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

References

This product has been referenced in:
  • Prieur A  et al. p53 and p16(INK4A) independent induction of senescence by chromatin-dependent alteration of S-phase progression. Nat Commun 2:473 (2011). Read more (PubMed: 21915115) »

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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