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Validated using a knockout cell line

Anti-CBX1 / HP1 beta antibody [MAC353] (ab10811)

Reviews (2)Q&A (3)References (13)
Western blot - Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade (ab10811)
  • Immunocytochemistry/ Immunofluorescence - Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade (ab10811)
  • Immunocytochemistry/ Immunofluorescence - Anti-CBX1 / HP1 beta antibody [MAC353] (ab10811)

Key features and details

  • Rat monoclonal [MAC353] to CBX1 / HP1 beta
  • Suitable for: WB, ICC/IF, IP, ELISA, IHC-Fr, ChIP
  • Knockout validated
  • Reacts with: Mouse, Human
  • Isotype: IgG2b

Overview

  • Product name

    Anti-CBX1 / HP1 beta antibody [MAC353]
    See all CBX1 / HP1 beta primary antibodies

  • Description

    Rat monoclonal [MAC353] to CBX1 / HP1 beta

  • Host species

    Rat

  • Specificity

    Ab10811 recognises the M31 molecule in mouse and the homologous HP1 HS beta molecule in man.

  • Tested applications

    Suitable for: WB, ICC/IF, IP, ELISA, IHC-Fr, ChIPmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Fusion protein corresponding to Mouse CBX1/ HP1 beta (C terminal).
    Database link: P83917

  • Positive control

    • Murine and human nuclear extracts.

  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

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: 0.09% Sodium azide
    Constituent: 5% BSA
  • Concentration information loading...

  • Purity

    Tissue culture supernatant

  • Clonality

    Monoclonal

  • Clone number

    MAC353

  • Isotype

    IgG2b

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab10811 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)
Use at an assay dependent concentration. Detects a band of approximately 26 kDa (predicted molecular weight: 22.2 kDa).
ICC/IF (1)
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.
ChIP
Use at an assay dependent concentration.
Notes
WB
Use at an assay dependent concentration. Detects a band of approximately 26 kDa (predicted molecular weight: 22.2 kDa).
ICC/IF
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.
ChIP
Use at an assay dependent concentration.

Target

  • Function

    Component of heterochromatin. Recognizes and binds histone H3 tails methylated at 'Lys-9', leading to epigenetic repression. Interaction with lamin B receptor (LBR) can contribute to the association of the heterochromatin with the inner nuclear membrane.

  • Tissue specificity

    Expressed in all adult and embryonic tissues.

  • Sequence similarities

    Contains 2 chromo domains.

  • Post-translational
    modifications

    Not phosphorylated.
    Ubiquitinated.

  • Cellular localization

    Nucleus. Unassociated with chromosomes during mitosis.
  • Information by UniProt

  • Database links

    • Alternative names

      • CBX 1 antibody
      • CBX antibody
      • Cbx1 antibody
      • CBX1_HUMAN antibody
      • Chromobox 1 antibody
      • Chromobox homolog 1 (HP1 beta homolog Drosophila ) antibody
      • chromobox homolog 1 antibody
      • Chromobox protein homolog 1 antibody
      • Drosophila HP1 beta antibody
      • Heterochromatin protein 1 beta antibody
      • Heterochromatin protein 1 homolog beta antibody
      • Heterochromatin protein p25 antibody
      • heterochromatin protein p25 beta antibody
      • HP1 beta antibody
      • HP1 beta homolog antibody
      • HP1 beta homolog Drosophila antibody
      • HP1, Drosophila. homolog of, beta antibody
      • HP1beta antibody
      • HP1Hs beta antibody
      • HP1Hsbeta antibody
      • Human heterochromatin protein p25 mRNA complete cds antibody
      • M31 antibody
      • MOD 1 antibody
      • MOD1 antibody
      • Modifier 1 protein antibody
      • p25beta antibody
      see all

    Images

    • Western blot - Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade (ab10811)
      Western blot - Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade (ab10811)

      Lane 1: Wild-type HAP1 cell lysate (40 µg)
      Lane 2: CBX1 knockout HAP1 cell lysate (40 µg)
      Lane 3: MCF7 cell lysate (40 µg)
      Lane 4: A431 cell lysate (40 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab10811 observed at 26 kDa. Red - loading control, ab181602, observed at 37 kDa.

      ab10811 was shown to specifically react with CBX1 / HP1 beta when CBX1 / HP1 beta knockout samples were used. Wild-type and CBX1 / HP1 beta knockout samples were subjected to SDS-PAGE. Ab10811 and ab181602 (loading control to GAPDH) were diluted at 1/500 and 1/10000 dilution respectively and incubated overnight at 4C. Blots were developed with Goat anti-Rat IgG H&L (IRDye® 800CW) (ab253031) preadsorbed and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777)secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade (ab10811)
      Immunocytochemistry/ Immunofluorescence - Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade (ab10811)This image is courtesy of Scott Slattery and Mike Mancini
      HeLA cells were stained with ab10811 in panel one. In panel two they were stained with ab10811 (green), DAPI (blue) and SH-CREST (red), which stains the centromeres. Fix 30 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with 1 mg/ml Na borohydride or 100 mM ammonium chloride in PEM. Permeablize 30 minutes with 0.5% TX-100 in PEM. Block 30 minutes in 5% milk in TBST. Primary antibody incubated overnight at 4oC diluted 1/100 in 5% milk in TBST. Secondary antibody incubated 1 hour at RT diluted in 5% milk in TBST. Post-fix 20 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with ammonium chloride in PEM. Counterstain with DAPI in TBST. Mount with ProLong Gold antifade reagent from Invitrogen. Notes: Ample washing between each step. TBST = Tris buffered saline + 0.1% Tween. PEM = 80 mM K-PIPES, pH 6.8, 5 mM EGTA, 2 mM MgCl2.
    • Immunocytochemistry/ Immunofluorescence - Anti-CBX1 / HP1 beta antibody [MAC353] (ab10811)
      Immunocytochemistry/ Immunofluorescence - Anti-CBX1 / HP1 beta antibody [MAC353] (ab10811)Image from Talasz H et al., J Biol Chem. 2005 Nov 18;280(46):38814-22. Epub 2005 Sep 15. Fig 1.; doi: 10.1074/jbc.M505563200; November 18, 2005 The Journal of Biological Chemistry, 280, 38814-38822.
      Immunofluorescence analysis of mouse erythroleukemia cell nuclei, staining CBX1 / HP1 beta in heterochromatin, with ab10811.

      Cells were fixed with paraformaldehyde, permeabilized using Triton X-100 and blocked for 30 min with 2.5% BSA. Cells were incubated with primary antibody (1/50) before incubating with a Cy3-conjugated goat anti-rat IgG to detect staining.

    References (13)

    Publishing research using ab10811? Please let us know so that we can cite the reference in this datasheet.

    ab10811 has been referenced in 13 publications.

    • Lamb KN  et al. Discovery of Potent Peptidomimetic Antagonists for Heterochromatin Protein 1 Family Proteins. ACS Omega 7:716-732 (2022). PubMed: 35036738
    • Otake K  et al. CENP-B creates alternative epigenetic chromatin states permissive for CENP-A or heterochromatin assembly. J Cell Sci 133:N/A (2020). PubMed: 32661090
    • Yang G  et al. The histone H3K9 methyltransferase SUV39H links SIRT1 repression to myocardial infarction. Nat Commun 8:14941 (2017). PubMed: 28361889
    • Himeda CL  et al. CRISPR/dCas9-mediated Transcriptional Inhibition Ameliorates the Epigenetic Dysregulation at D4Z4 and Represses DUX4-fl in FSH Muscular Dystrophy. Mol Ther 24:527-35 (2016). ChIP ; Human . PubMed: 26527377
    • Liu J  et al. HP1a mediates defective heterochromatin repair and accelerates senescence in Zmpste24-deficient cells. Cell Cycle 13:1237-47 (2014). PubMed: 24584199
    View all Publications for this product

    Customer reviews and Q&As

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

    1-5 of 5 Abreviews or Q&A

    Western blot abreview for Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (RAW264.7 cells)
    Gel Running Conditions
    Reduced Denaturing (14 %)
    Loading amount
    80 µg
    Specification
    RAW264.7 cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Magali Boissiere

    Verified customer

    Submitted Feb 22 2017

    Immunocytochemistry/ Immunofluorescence abreview for Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade

     Average
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Colorectal Cancer Cells)
    Specification
    Colorectal Cancer Cells
    Fixative
    Formaldehyde
    Permeabilization
    Yes - triton x-100
    Blocking step
    Serum as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jan 21 2010

    Question

    Hi，
    I ordered a primary antibody (Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade (ab10811)) and I plan to use this Ab for ChIP. But the concentration of this Ab is unknown. I wanna ask whether you have any reference data for ChIP using this Ab.
    Thank you very much!


    --

    Read More
    Answer

    Thank you for contacting us.


    For ChIP, we recommend to optimise the working concentration according to sample and experiment. At the starting point, you could try using 1:50 dilution.


    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

    Question

    I just wanted to know if this antibody CBX1 MAC 353 (ab10811) is only detecting CBX1 without phosphorylated sites on Thr51 or with phosphorylated sites on Thr51? Or is it just the whole amount of CBX1-protein?

    Read More
    Answer

    Thank you for contacting us with your questions. Since the immunogen of the antibody was taken from the C-terminus, and threonine 51 is much closer to the N-terminus, the antibody will detect the total CBX1 protein. The antibody will recognize CBX1 whether or not it is phosphorylated at threonine 51. I hope this information is helpful, but please let me know if you have any further questions.

    Read More

    Question

    I received antibody, and thanks for the pretty mug cup. I like to know the concentration.What's the recommended dilution for western and immunofluorescence?

    Read More
    Answer

    I'm glad you like the mug! We are unable to provide a concentration for ab10811 as it is in an unpurified form, tissue culture supernatant, and we cannot guarantee the concentration for unpurified material. I recommend starting at a dilution of 1:1000 and optimizing from there.

    Read More

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