Validated using a knockout cell lineRecombinantRabMAb

Recombinant HRP Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab197721)

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Western blot - HRP Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab197721)
  • Western blot - HRP Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab197721)
  • HRP Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab197721)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • HRP Rabbit monoclonal [EPR5777] to HP1 alpha - Heterochromatin marker
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Human
  • Conjugation: HRP

Overview

  • Product name

    HRP Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker
    See all HP1 alpha primary antibodies

  • Description

    HRP Rabbit monoclonal [EPR5777] to HP1 alpha - Heterochromatin marker

  • Host species

    Rabbit

  • Conjugation

    HRP

  • Tested applications

    Suitable for: WBmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat

  • Immunogen

    Synthetic peptide within Human HP1 alpha aa 150-250. The exact sequence is proprietary.

  • Positive control

    • HeLa, SH-SY5Y, MCF7, A431 cell lysates and Human tonsil tissue

  • General notes

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab197721 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 1/5000. Detects a band of approximately 28 kDa (predicted molecular weight: 22 kDa).

Target

  • Function

    Component of heterochromatin that recognizes and binds histone H3 tails methylated at 'Lys-9' (H3K9me), leading to epigenetic repression. In contrast, it is excluded from chromatin when 'Tyr-41' of histone H3 is phosphorylated (H3Y41ph). Can interact with lamin-B receptor (LBR). This interaction can contribute to the association of the heterochromatin with the inner nuclear membrane. Involved in the formation of functional kinetochore through interaction with MIS12 complex proteins.

  • Sequence similarities

    Contains 2 chromo domains.

  • Post-translational
    modifications

    Phosphorylation of HP1 and LBR may be responsible for some of the alterations in chromatin organization and nuclear structure which occur at various times during the cell cycle (By similarity). Phosphorylated during interphase and possibly hyper-phosphorylated during mitosis.
    Ubiquitinated.

  • Cellular localization

    Nucleus. Chromosome. Chromosome > centromere. Component of centromeric and pericentromeric heterochromatin. Associates with chromosomes during mitosis. Associates specifically with chromatin during metaphase and anaphase.
  • Information by UniProt

  • Database links

    • Alternative names

      • Antigen p25 antibody
      • CBX5 antibody
      • CBX5_HUMAN antibody
      • CG8409 antibody
      • Chromobox 5 antibody
      • Chromobox homolog 5 (HP1 alpha homolog, Drosophila) antibody
      • Chromobox homolog 5 antibody
      • Chromobox protein homolog 5 antibody
      • Epididymis luminal protein 25 antibody
      • HEL25 antibody
      • Heterochromatin protein 1 alpha antibody
      • Heterochromatin protein 1 antibody
      • Heterochromatin protein 1 homolog alpha antibody
      • HP1 alpha antibody
      • HP1 alpha homolog antibody
      • HP1 antibody
      • HP1A antibody
      • HP1Hs alpha antibody
      • Su(var)205 antibody
      see all

    Images

    • Western blot - HRP Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab197721)
      Western blot - HRP Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab197721)
      All lanes : HRP Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab197721) at 1/5000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : CBX5 (HP1 alpha) knockout HAP1 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 22 kDa


      Exposure time: 20 minutes


      ab197721 was shown to specifically react with HP1 alpha in wild-type HAP1 cells as signal was lost in CBX5 (HP1 alpha) knockout cells. Wild-type and CBX5 (HP1 alpha) knockout samples were subjected to SDS-PAGE. Ab197721 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/1000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

    • Western blot - HRP Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab197721)
      Western blot - HRP Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab197721)
      All lanes : HRP Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab197721) at 1/5000 dilution

      Lane 1 : SH-SY5Y (Human neuroblastoma cell line) Whole Cell Lysate at 10 µg
      Lane 2 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg
      Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
      Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
      Lane 5 : Hap1 WT at 20 µg
      Lane 6 : HP1 alpha knockout HAP1 cell lysate at 20 µg

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 22 kDa
      Observed band size: 28 kDa
      why is the actual band size different from the predicted?


      Exposure time: 16 minutes


      This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab197721 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

    • HRP Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab197721)
      HRP Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab197721)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab197721? Please let us know so that we can cite the reference in this datasheet.

    ab197721 has not yet been referenced specifically in any publications.

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