Overview

  • Product name

    Anti-Hho1 antibody
  • Description

    Rabbit polyclonal to Hho1
  • Host species

    Rabbit
  • Specificity

    This antibody recognizes linker histone from S. Cerevisiae.
  • Tested applications

    Suitable for: ChIP, CHIPseq, WBmore details
  • Species reactivity

    Reacts with: Saccharomyces cerevisiae
  • Immunogen

    Recombinant full length protein purified from E. coli.

Properties

Applications

Our Abpromise guarantee covers the use of ab71833 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
ChIP Use at an assay dependent concentration. PubMed: 22689825
CHIPseq Use at an assay dependent concentration. PubMed: 22689825
WB 1/1000. Predicted molecular weight: 28 kDa.

Target

  • Relevance

    Hho1 could act as an H1-type linker histone. It has been shown to bind DNA.
  • Cellular localization

    Nuclear
  • Database links

    • Alternative names

      • Hho1p antibody
      • Histone H1, a linker histone required for nucleosome packaging at restricted sites antibody
      • LPI17C antibody
      • Not required for telomeric silencing, basal transcriptional repression, or efficient sporulation antibody
      • Suppresses DNA repair involving homologous recombination antibody
      • YPL127C antibody
      see all

    Images

    • Lane
      1: 50 ng recombinant Hho1.
      2: WCE from Hho1 knockout yeast expressing deletion construct A.
      3: WCE from Hho1 knockout yeast expressing deletion construct B.
      4: WCE from Hho1 knockout yeast expressing deletion construct C.
      5: WCE from Hho1 knockout yeast expressing deletion construct D.
      6: WCE from Hho1 knockout yeast expressing deletion construct E.
      7: WCE from hho1 knockout yeast expressing full-length Hho1(F).
      8: WCE from hho1 knockout yeast carrying an empty vector.
      9: WCE from hho1 knockout yeast.
      10: WCE from wild-type yeast.
    • All lanes : Anti-Hho1 antibody (ab71833) at 1/1000 dilution

      Lane 1 : Wild type S. cerevisiae whole cell lysate
      Lane 2 : Hho1 knock-out S. cerevisiae whole cell lysate
      Lane 3 : S. cerevisiae whole cell lysate expressing Hho1 with tag

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : HRP-conjugated goat anti-rabbit polyclonal IgG at 1/5000 dilution

      Predicted band size: 28 kDa
      Observed band size: 35 kDa
      why is the actual band size different from the predicted?

      See Abreview

    References

    This product has been referenced in:

    • Lawrence MBD  et al. Histone Acetylation, Not Stoichiometry, Regulates Linker Histone Binding in Saccharomyces cerevisiae. Genetics 207:347-355 (2017). Read more (PubMed: 28739661) »
    • Brush GS Evidence that histone H1 is dispensable for proper meiotic recombination in budding yeast. BMC Res Notes 8:275 (2015). Read more (PubMed: 26122007) »
    See all 4 Publications for this product

    Customer reviews and Q&As

    1-5 of 5 Abreviews or Q&A

    Application
    Western blot
    Sample
    Saccharomyces cerevisiae Cell lysate - whole cell (cycling haploid cells)
    Loading amount
    15 µg
    Specification
    cycling haploid cells
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Jan 23 2013

    Answer

    Thank you for contacting us.

    I have checked with the lab, butunfortunately, they are unable to provide the antibody in purified, glycerol-free form.

    I am sorry to not have more helpful information, but hope my recent email gave you some alternative options to consider. Please let me know if I can be of further help.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Answer

    Thank you for your email.

    The antibody ab71833 comes as whole antiserum and is therefore not purified. The antibody concentration has not been determined as it would not be reflective of the concentration of the specific antibody in the serum. In general, if the total antibody concentration in serum is 10 mg/ml, then only a max. of 10%(or 1 mg/ml) would bespecific antibody. These are general estimates.

    For conjugation purposes, you would need to purify the antibody from other proteins that could also get conjugated and interfere with the reaction. We carry purification kits for that purpose:

    https://www.abcam.com/index.html?pageconfig=resource&rid=13489

    To purify serum or ascites fluid - up to 20 mg of antibody

    Abcam's antibody serum purification kits are prepared by coupling highly purified protein A or G to agarose beads and can therefore be used to purify IgG fractions from both serum and ascites fluid. The antibody is captured on the agarose beads and unwanted substances are removed by a simple wash procedure. The purified product is then eluted and neutralized. Antibody serum purification kits can purify up to 20 mg of antibody. The volume of sample required will depend on the host species and will be mentioned on each datasheet.

    Protein G has a high affinity for the Fc regions of IgG molecules from a variety of species (e.g. sheep, rat or goat). Protein A resin has a high affinity for the Fc regions of rabbit IgG molecules

    Protein A Antibody serum purification kit (1 to 3 purifications): https://www.abcam.com/Antibody-Serum-Purification-Kit-Protein-A-ab109209.html
    Protein G Antibody serum purification kit (1 to 3 purifications): https://www.abcam.com/Serum-Antibody-Purification-Kit-Protein-G-ab128751.html



    I would recommend ab109209 since the antibody is a rabbit polyclonal.

    Alternatively, I can check with the lab if the antibody could be purified, but this would be a custom preparation and would require an order of at least 10 vials. I'll let you know what I find out.

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

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Answer

    I now have further information to share with the customer in regards to the protocol used to perform the western blotting experiment presented on the datasheet ofthe Anti-Hho1 antibody (ab71833).


    Theloading buffer used was a standard SDSloading buffer. We have a recipie for this in our protocol book which can be accessed from the following link:


    https://docs.abcam.com/pdf/misc/abcam-protocols-book-2010.pdf


    No protease inhibitors were used in the lysis as TCA was used to lyse the cells and therefore they were not required.


    Blocking of the membrane was performed using 5% Milk in TBS-T for 1 hour at room temperature. The primary and secondary antibody was diluted in TBS-T.


    I hope this information has been of help. If you require any further information please do not hesitate to ask.

    Read More

    Answer

    Thank you for contacting us.


    The Western blot presented on the datasheet ofAnti-Hho1 antibody (ab71833) wasperformed using extracts prepared by glass bead disruption into 20% TCA (the cell pellet was prepared from a 10 ml mid-log culture of the wildetype or hho1 deletion strain). After lysing, the precipitatewas washed twice in 100% acetone, then air dried and resuspended in 1x SDS loading buffer (usually about 100 ul for cells from 10 ml of culture). Thiswas then boiled, and about 20 ul loaded onto the gel. The ab71833 was then used in a 1/1000 dilution with incubation for 1 hour at room temperature. Ananti-rabbit secondary antibody was then used for detection.


    I hope this information has been of help. If you require any further information please do not hesitate to contact us again.

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

    Sign up