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Epigenetics and Nuclear Signaling Histones Variants
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Validated using a knockout cell line

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

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Western blot - Anti-Histone H1.0 antibody [27] (ab11080)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [27] (ab11080)
  • Western blot - Anti-Histone H1.0 antibody [27] (ab11080)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [27] (ab11080)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [27] (ab11080)
  • Flow Cytometry - Anti-Histone H1.0 antibody [27] (ab11080)

Key features and details

  • Mouse monoclonal [27] to Histone H1.0
  • Suitable for: WB, Flow Cyt, IHC-P
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

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Overview

  • Product name

    Anti-Histone H1.0 antibody [27]
    See all Histone H1.0 primary antibodies
  • Description

    Mouse monoclonal [27] to Histone H1.0
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, Flow Cyt, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Cow, Xenopus laevis, VertebrataDoes not react with: Bird
  • Immunogen

    Full length native protein (purified) corresponding to Cow Histone H1.0.

  • Epitope

    This 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

    • WB: A431, MCF7 and HeLa cell lysates; Histone H1.0 Human Recombinant Protein. IHC-P: Human colon and pancreas adenocarcinoma tissues.
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

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

    Preservative: 0.02% Sodium azide
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...
  • Purity

    IgG fraction
  • Clonality

    Monoclonal
  • Clone number

    27
  • Myeloma

    NS1/1-Ag4-1
  • Isotype

    IgG1
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Histones
    • Variants
    • Epigenetics and Nuclear Signaling
    • Histones
    • H1
    • Unmodified

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • Recombinant Protein

    • Recombinant Human Histone H1.0 protein (ab158626)

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
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.
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.

Target

  • Function

    Histones 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 similarities

    Belongs 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 localization

    Nucleus. Chromosome. The RNA edited version has been localized to nuclear speckles. During mitosis, it appears in the vicinity of condensed chromosomes.
  • Target information above from: UniProt accession P07305 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3005 Human
    • Entrez Gene: 14958 Mouse
    • Entrez Gene: 24437 Rat
    • Omim: 142708 Human
    • SwissProt: Q0IIJ2 Cow
    • SwissProt: P07305 Human
    • SwissProt: P10922 Mouse
    • SwissProt: P43278 Rat
    • Unigene: 745024 Human
    • Unigene: 24350 Mouse
    • Unigene: 3129 Rat
    see all
  • 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

Images

  • Western blot - Anti-Histone H1.0 antibody [27] (ab11080)
    Western blot - Anti-Histone H1.0 antibody [27] (ab11080)
    All lanes : Anti-Histone H1.0 antibody [27] (ab11080) at 1/500 dilution

    Lane 1 : Wild-type A431 cell lysate
    Lane 2 : H1F0 knockout A431 cell lysate
    Lane 3 : MCF7 cell lysate

    Lysates/proteins at 40 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 20 kDa
    Observed band size: 30 kDa
    why is the actual band size different from the predicted?



    Lanes 1 - 3: Merged signal (red and green). Green - ab11080 observed at 30 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa.

    ab11080 was shown to react with Histone H1.0 in wild-type A431 cells in Western blot with loss of signal observed in H1F0 knockout sample. Wild-type A431 and H1F0 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab11080 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [27] (ab11080)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [27] (ab11080)

    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

  • Western blot - Anti-Histone H1.0 antibody [27] (ab11080)
    Western blot - Anti-Histone H1.0 antibody [27] (ab11080)
    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
    All lanes : 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
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [27] (ab11080)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [27] (ab11080)

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [27] (ab11080)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [27] (ab11080)

    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.

  • Flow Cytometry - Anti-Histone H1.0 antibody [27] (ab11080)
    Flow Cytometry - Anti-Histone H1.0 antibody [27] (ab11080)
    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.

Protocols

  • Mouse on Mouse staining protocol

Click here to view the general protocols

Datasheets and documents

    • Datasheet
  • References (9)

    Publishing research using ab11080? Please let us know so that we can cite the reference in this datasheet.

    ab11080 has been referenced in 9 publications.

    • 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 . PubMed: 27611779
    • Fu G  et al. Mouse oocytes and early embryos express multiple histone H1 subtypes. Biol Reprod 68:1569-76 (2003). PubMed: 12606334
    • Khochbin S Histone H1 diversity: bridging regulatory signals to linker histone function. Gene 271:1-12 (2001). PubMed: 11410360
    • Adenot PG  et al. Somatic linker histone H1 is present throughout mouse embryogenesis and is not replaced by variant H1 degrees. J Cell Sci 113 ( Pt 16):2897-907 (2000). PubMed: 10910774
    • Grunwald D  et al. In situ analysis of chromatin proteins during development and cell differentiation using flow cytometry. Methods Mol Biol 119:443-54 (1999). PubMed: 10804531
    • Gorka C  et al. Differential recognition of histone H10 by monoclonal antibodies during cell differentiation and the arrest of cell proliferation. J Biol Chem 273:1208-15 (1998). PubMed: 9422788
    • Gorka C  et al. Variation of H1(0) content throughout the cell cycle in regenerating rat liver. Exp Cell Res 217:528-33 (1995). PubMed: 7698253
    • Grunwald D  et al. Accumulation of histone H1(0) during early Xenopus laevis development. Exp Cell Res 218:586-95 (1995). PubMed: 7796895
    • Seigneurin D  et al. Developmentally regulated chromatin acetylation and histone H1(0) accumulation. Int J Dev Biol 39:597-603 (1995). PubMed: 8619958

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