Overview

  • Product nameAnti-Histone H1.0 antibody [27]
    See all Histone H1.0 primary antibodies
  • Description
    Mouse monoclonal [27] to Histone H1.0
  • SpecificityThis clone only recognises Histone H1.0 in differentiating cells, whereas clone 34 recognises H1.0 constitutively. This antibody recognises a region of the protein at the N-terminal part of the globular domain. It will not recognise H1.0 in chromatin, whereas clone 34 will do.
  • Tested applicationsSuitable for: ICC/IF, WB, IHC-Fr, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, Xenopus laevis
    Predicted to work with: all other VertebratesDoes not react with: Bird
  • Immunogen

    Ox Liver Histone H1.0.

  • EpitopeThis antibody recognises an epitope within aa24-30. Proline 26, which is responsible for a bend in this region, plays an important role in the recognition. See Gorka et al. 1998 for more information.
  • Positive control
    • Murine erythroleukemia (MEL) cells were used in part of the original characterisation of this antibody. Purified histones can also be used as a positive control. In Flow Cytometry, this antibody gave a positive signal in methanol fixed/Tween permeabilised HeLa cells. This antibody gave a positive result in IHC in the following FFPE tissue: Human pancreas adenocarcinoma.

Properties

Applications

Our Abpromise guarantee covers the use of ab11080 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
ICC/IF Use at an assay dependent concentration.
WB 1/500. Detects a band of approximately 30 kDa (predicted molecular weight: 20 kDa). Linker histones run at about 30kD even though the predicted size is about 20kD.
IHC-Fr Use at an assay dependent concentration. Formalin fixed .
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

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.
EMSA Use at an assay dependent concentration.

Target

  • FunctionHistones H1 are necessary for the condensation of nucleosome chains into higher-order structures. The H1F0 histones are found in cells that are in terminal stages of differentiation or that have low rates of cell division.
  • Sequence similaritiesBelongs to the histone H1/H5 family.
    Contains 1 H15 (linker histone H1/H5 globular) domain.
  • Post-translational
    modifications
    Phosphorylated on Ser-17 in RNA edited version.
  • Cellular localizationNucleus. Chromosome. The RNA edited version has been localized to nuclear speckles. During mitosis, it appears in the vicinity of condensed chromosomes.
  • Information by UniProt
  • Database links
  • Alternative names
    • H1 histone family member 0 antibody
    • H1(0) antibody
    • H10 antibody
    • H10_HUMAN antibody
    • h1f0 antibody
    • H1FV antibody
    • Histone H1'' antibody
    • Histone H1(0) antibody
    • Histone H1.0 antibody
    • Histone H10 antibody
    • Histone H5 antibody
    • MGC5241 antibody
    • N-terminally processed antibody
    see all

Anti-Histone H1.0 antibody [27] images

  • IHC image of Histone H1 staining in a section of formalin-fixed paraffin-embedded [human normal colon]*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was then incubated with ab11080, 1/1000 dilution, for 15 mins at room temperature. A goat anti-mouse biotinylated secondary antibody (ab6788, 1/1000 dilution), was used to detect the primary, and visualized using an HRP conjugated ABC system. Streptavidin HRP was used, ab7403 at a 1/10000 dilution. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10min at room temperature. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

    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 Histone H1.0 staining in human colon formalin fixed paraffin embedded tissue section*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab11080, 7.5µg/ml overnight at +4°C. An HRP-conjugated secondary (ab97240, 1/2000 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.

    The inset negative control image is secondary-only at 1/500 dilution.

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

  • IHC image of Histone H1.0 staining in Human pancreas adenocarcinoma 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 ab11080, 5µg/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 H1.0 antibody [27] (ab11080) at 1/500 dilution

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 3 : Histone H1.0 Human Recombinant Protein

    Lysates/proteins at 30 µg per lane.

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

    Performed under reducing conditions.

    Predicted band size : 20 kDa
    Observed band size : 30 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 46 kDa,65 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 1 minute
  • ICC/IF image of ab11080 stained MCF7 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 (ab11080, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879 Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/250 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.

  • Overlay histogram showing HeLa cells stained with ab11080 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab11080, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References for Anti-Histone H1.0 antibody [27] (ab11080)

This product has been referenced in:
  • Yan X  et al. CaMKII-Mediated CREB Phosphorylation Is Involved in Ca2+-Induced BDNF mRNA Transcription and Neurite Outgrowth Promoted by Electrical Stimulation. PLoS One 11:e0162784 (2016). WB ; Rat . Read more (PubMed: 27611779) »
  • Fu G  et al. Mouse oocytes and early embryos express multiple histone H1 subtypes. Biol Reprod 68:1569-76 (2003). Read more (PubMed: 12606334) »

See all 9 Publications for this product

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