Validated using a knockout cell line
Recombinant
RabMAb

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

Overview

  • Product name

    Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (HRP)
    See all HP1 alpha primary antibodies
  • Description

    Rabbit monoclonal [EPR5777] to HP1 alpha - Heterochromatin marker (HRP)
  • Host species

    Rabbit
  • Conjugation

    HRP
  • Tested applications

    Suitable for: IHC-P, 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

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

    This product is a recombinant rabbit monoclonal antibody.

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
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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

  • All lanes : Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (HRP) (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.

  • All lanes : Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (HRP) (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.

References

ab197721 has not yet been referenced specifically in any publications.

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