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

Overview

  • Product name
    Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade
    See all CBX1 / HP1 beta primary antibodies
  • Description
    Rat monoclonal [MAC353] to CBX1 / HP1 beta - ChIP Grade
  • Specificity
    Ab10811 recognises the M31 molecule in mouse and the homologous HP1 HS beta molecule in man.
  • Tested applications
    Suitable for: IHC-Fr, ELISA, WB, ICC/IF, ChIP, IPmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Fusion protein corresponding to Human CBX1/ HP1 beta (C terminal).

  • Positive control
    • Murine and human nuclear extracts.

Properties

Applications

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
IHC-Fr 1/1 - 1/50.
ELISA Use at an assay dependent dilution.
WB Use at an assay dependent dilution. Detects a band of approximately 26 kDa (predicted molecular weight: 22.2 kDa).
ICC/IF Use at an assay dependent dilution.
ChIP Use at an assay dependent concentration.
IP Use at an assay dependent dilution.

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

Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade images



  • Predicted band size : 22.2 kDa

    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-Mouse IgG H&L (IRDye® 800CW)  preadsorbed (ab216772) 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.

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

This product has been referenced in:
  • Yang G  et al. The histone H3K9 methyltransferase SUV39H links SIRT1 repression to myocardial infarction. Nat Commun 8:14941 (2017). Read more (PubMed: 28361889) »
  • Vaskova EA  et al. Dynamics of the two heterochromatin types during imprinted X chromosome inactivation in vole Microtus levis. PLoS One 9:e88256 (2014). ICC/IF . Read more (PubMed: 24505450) »

See all 7 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Magali Boissiere

Verified customer

Submitted Feb 22 2017

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 ...

Read More

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 ...

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Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Jan 21 2010

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 ...

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