Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Histone H1.0 antibody [EPR6537] (ab125027)

Submit a review Q&A (2)References (10)

Overview

  • Product name

    Anti-Histone H1.0 antibody [EPR6537]
    See all Histone H1.0 primary antibodies

  • Description

    Rabbit monoclonal [EPR6537] to Histone H1.0

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human Histone H1.0 aa 150 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P07305

  • Positive control

    • WB: A431 and BxPC3 cell lysates, and fetal human lung lysates IHC-P: Human kidney, pancreas, and thyroid, rat kidney, mouse liver tissues. ICC/IF: BxPC-3 and A431 cells.

  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab125027 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 1/1000 - 1/10000. Detects a band of approximately 32 kDa (predicted molecular weight: 21 kDa).
IHC-P 1/100 - 1/400. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100 - 1/250.

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.
  • 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

    Images

    • Western blot - Anti-Histone H1.0 antibody [EPR6537] (ab125027)
      Western blot - Anti-Histone H1.0 antibody [EPR6537] (ab125027)
      All lanes : Anti-Histone H1.0 antibody [EPR6537] (ab125027) at 1/1000 dilution

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

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 21 kDa
      Observed band size: 33 kDa
      why is the actual band size different from the predicted?



      Lanes 1 - 2: Merged signal (red and green). Green - ab125027 observed at 33 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

      ab125027 was shown to react with H1F0 in A431 wild-type cells in Western blot. Loss of signal was observed when H1F0 knockout sample was used. A431 wild-type 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 ab125027 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-Histone H1.0 antibody [EPR6537] (ab125027)
      Western blot - Anti-Histone H1.0 antibody [EPR6537] (ab125027)
      All lanes : Anti-Histone H1.0 antibody [EPR6537] (ab125027) at 10 µg (purified)

      Lane 1 : human fetal lung lysate
      Lane 2 : BxPC-3 lysate
      Lane 3 : A431 lysate
      Lane 4 : human fetal kidney lysate

      Secondary
      All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 21 kDa
      Observed band size: 32 kDa why is the actual band size different from the predicted?



      Blocking buffer: 5% NFDM/TBST
      Dilution buffer: 5% NFDM/TBST

    • Immunocytochemistry/ Immunofluorescence - Anti-Histone H1.0 antibody [EPR6537] (ab125027)
      Immunocytochemistry/ Immunofluorescence - Anti-Histone H1.0 antibody [EPR6537] (ab125027)

      Immunofluorescence staining of BxPC-3 cells with purified ab125027 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 100% methanol and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab125027 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [EPR6537] (ab125027)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [EPR6537] (ab125027)
      Immunohistochemical staining of paraffin embedded rat kidney with purified ab125027 at a working dilution of 1/400. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [EPR6537] (ab125027)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [EPR6537] (ab125027)
      Immunohistochemical staining of paraffin embedded mouse liver with purified ab125027 at a working dilution of 1/400. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [EPR6537] (ab125027)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [EPR6537] (ab125027)
      Immunohistochemical staining of paraffin embedded human thyroid carcinoma with purified ab125027 at a working dilution of 1/400. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [EPR6537] (ab125027)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [EPR6537] (ab125027)
      Immunohistochemical staining of paraffin embedded human pancreas with purified ab125027 at a working dilution of 1/400. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
    • Western blot - Anti-Histone H1.0 antibody [EPR6537] (ab125027)
      Western blot - Anti-Histone H1.0 antibody [EPR6537] (ab125027)
      All lanes : Anti-Histone H1.0 antibody [EPR6537] (ab125027) at 1/1000 dilution (unpurified)

      Lane 1 : BxPC3 lysates
      Lane 2 : Human fetal lung lysates

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti rabbit at 1/2000 dilution

      Predicted band size: 21 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [EPR6537] (ab125027)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [EPR6537] (ab125027)

      Unpurified ab125027, at a 1/100 dilution, staining Histone H1 in paraffin embedded Human kidney tissue by Immunohistochemistry.

    • Immunocytochemistry/ Immunofluorescence - Anti-Histone H1.0 antibody [EPR6537] (ab125027)
      Immunocytochemistry/ Immunofluorescence - Anti-Histone H1.0 antibody [EPR6537] (ab125027)

      Unpurified ab125027, at a 1/100 dilution, staining Histone H1 in A431 cells by Immunofluorescence.

    References

    This product has been referenced in:

    • Steele NG  et al. An Organoid-Based Preclinical Model of Human Gastric Cancer. Cell Mol Gastroenterol Hepatol 7:161-184 (2019). Read more (PubMed: 30522949) »
    • Tao L  et al. Polypharmacological Profiles Underlying the Antitumor Property of Salvia miltiorrhiza Root (Danshen) Interfering with NOX-Dependent Neutrophil Extracellular Traps. Oxid Med Cell Longev 2018:4908328 (2018). Read more (PubMed: 30210653) »
    See all 10 Publications for this product

    Publishing research using ab125027? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-2 of 2 Abreviews or Q&A

    Question

    Thank you for your response. Histone H1.2, 1.3 or 1.4 are about 21-22KD. Which of your antibody recognize these three isoforms, especially H1.4?
    I also notice that you have Anti-Histone H1 (tri methyl K25) antibody (ab17347) which detects H1.4K25. But there is a post on your web site (question 2) states:"This antibody may not recognize methylated H1 but only the H3." Do you have any update for it? I am
    interested to purchase this antibody if it indeed can recognize H1.4K25(K26).

    Read More
    Answer

    Thank you for contacting us.

    H1.2 specific antibodies,
    ab4086; https://www.abcam.com/Histone-H1-2-antibody-ChIP-Grade-ab4086.html. (we however haven't tested the specificity to other forms)
    ab17677; https://www.abcam.com/Histone-H1-2-antibody-ab17677.html

    H1.3 abs
    ab24174; https://www.abcam.com/Histone-H1-3-antibody-ab24174.html
    H1.4
    ab105522; https://www.abcam.com/Histone-H1-4-antibody-ab105522.html
    ab116393; https://www.abcam.com/Histone-H1-4-antibody-ab116393.html

    The antibody ab17347 is for tri methylated H1, unfortunately it also cross react with methylated histone H3 and other isoform of H1. This antibody however will not recognize di and mono methylated histone H1. Is this the answer to your question? Let me know if you any further question about the specificity of these products.

    Read More

    Question

    Would you please help to verify the following questions regarding your
    anti-histone H1 anti bodies:
    Anti-Histone H1 antibody (ab87556): Detects a ˜32KD band;
    Anti-Histone H1 antibody (ab83058) : Detects a ˜24KD band;
    Anti-Histone H1 antibody [1415-1] - Carboxyterminal end (ab62884) :
    Detects a ˜17KD band;
    Anti-Histone H1 antibody [AE-4] (ab71594) : no WB image;
    Anti-Histone H1 antibody [EPR6537] (ab125027): WB ˜32KD
    1). Do they represent different modifications of H1 or different isofroms
    of H1, since the band sizes recognized are so diffenrent? (But MW of H1
    isoforms are around 22KD).
    2). Will above antibodies also cross-react with other histones like H3 or H4?
    Anti-Histone H1 antibody [0.T.60] (ab4269): Detects isoforms 0-5,
    ˜25-32KD; Why the sizes are different from the oredicted sizes, or due to
    modification or something else?
    In the end, we need an antibody to detect Histone H1 in human cell lines
    such as HEK293 and SH-SY5Y, which one do you recommend?
    Thank you!

    Read More
    Answer

    Thank you for contacting us.

    ab87556 was used with a sequence from H1FOO_HUMAN protein; http://www.uniprot.org/uniprot/Q8IZA3. According to swissprot the molecular weight is 35 kDa.

    ab83058; the sequence used is from P07305 http://www.uniprot.org/uniprot/P07305. The molecular weight is ˜ 21kDa.

    ab71594 was used with a sequence from H1FOO_HUMAN protein; http://www.uniprot.org/uniprot/Q8IZA3. According to swissprot the molecular weight is ˜ 35 kDa.

    ab125027 was used with a sequence from H1FOO_HUMAN protein; http://www.uniprot.org/uniprot/Q8IZA3. According to swissprot the molecular weight is ˜ 35 kDa.

    These antibodies are specific to human H1 protein only not the modifications. You can purchase any of these to detect H1 human protein.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

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