Overview

  • Product nameAnti-Histone H4 antibody
    See all Histone H4 primary antibodies
  • Description
    Rabbit polyclonal to Histone H4
  • Specificityab52178 detects endogenous levels of total Histone H4 protein
  • Tested applicationsSuitable 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

  • FormLiquid
  • Storage instructionsShipped 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 bufferPreservative: 0.02% Sodium Azide
    Constituents: 50% Glycerol, PBS (without Mg2+ and Ca2+), 150mM Sodium chloride, pH 7.4
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • Purification notesab52178 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.
  • ClonalityPolyclonal
  • IsotypeIgG
  • 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

Anti-Histone H4 antibody 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 for Anti-Histone H4 antibody (ab52178)

ab52178 has not yet been referenced specifically in any publications.

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