Overview

  • Product name
    Anti-CBX1 / HP1 beta antibody
    See all CBX1 / HP1 beta primary antibodies
  • Description
    Rabbit polyclonal to CBX1 / HP1 beta
  • Host species
    Rabbit
  • Specificity
    ab10478 recognises a band at 25 kDa, which is attributed to CBX1 / HP1 beta.
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Marsupial
    Predicted to work with: Chicken, Hamster, Drosophila melanogaster
  • Immunogen

    Synthetic peptide, conjugated to KLH, derived from within residues 150 to the C-terminus of Human HP1 beta.

    Read Abcam's proprietary immunogen policy .

  • Positive control
    • This antibody gave a positive signal in the following lysates: HeLa Whole Cell, HeLa Nuclear, A431 Whole Cell, MCF7 Whole Cell, HEK293 Whole Cell, PC12 Whole Cell

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: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab10478 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 1/500 - 1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 21 kDa).
IHC-P 1/100.
ICC/IF 1/200.
IP 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

  • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: CBX1 knockout  HAP1 whole cell lysate (20 µg)
    Lane 3: MCF7 whole cell lysate (20 µg)
    Lane 4: A431 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab10478 observed at 26 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab10478 was shown to recognize CBX1 when CBX1 knockout samples were used, along with additional cross-reactive bands. Wild-type and CBX1 knockout samples were subjected to SDS-PAGE. Ab10478 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-CBX1 / HP1 beta antibody (ab10478) at 1 µg/ml

    Lane 1 : HeLa whole cell lysate
    Lane 2 : HeLa nuclear lysate
    Lane 3 : A431 cell lysate
    Lane 4 : MCF7 cell lysate
    Lane 5 : HEK293 cell lysate
    Lane 6 : HeLa whole cell lysate with CBX1 / HP1 beta peptide (ab30861) at 1 µg/ml
    Lane 7 : HeLa nuclear lysate with CBX1 / HP1 beta peptide (ab30861) at 1 µg/ml
    Lane 8 : A431 lysate with CBX1 / HP1 beta peptide (ab30861) at 1 µg/ml
    Lane 9 : MCF7 cell lysate with CBX1 / HP1 beta peptide (ab30861) at 1 µg/ml
    Lane 10 : HEK293 cell lysate with CBX1 / HP1 beta peptide (ab30861) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

    Predicted band size: 21 kDa
    Observed band size: 25 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 37 kDa (possible cross reactivity), 47 kDa (possible cross reactivity)

  • CBX1 / HP1 beta was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to CBX1 / HP1 beta and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab10478.
    Secondary: Clean blot (HRP conjugate) at 1/1000 dilution.
    Band: 20ka: CBX1 / HP1 beta.
  • ab10478 at 1/200 staining human cervix cells by ICC/IF. The cells were fixed in 4% PFA for 15 minutes and permeabilized for 15 minutes in 0.5% Triton X100 in PBS, prior to incubation with the antibody fort 1 hour. A Cy2 ®  conjugated goat anti-rabbit antibody was used as the secondary.

    See Abreview

  • Image courtesy of Human Protein Atlas

    ab10478 staining CBX1 / HP1 in human testis, showing a distinct and strong staining pattern in ductus seminiferus cells. Paraffin embedded human skin tissue was incubated with ab10478 (1/400 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

    ab10478 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org

    .
  • Anti-CBX1 / HP1 beta antibody (ab10478) at 1 µg/ml + PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 21 kDa
    Observed band size: 27 kDa why is the actual band size different from the predicted?
    Additional bands at: 90 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 4 minutes

References

This product has been referenced in:
  • Yang YF  et al. CBX1 Indicates Poor Outcomes and Exerts Oncogenic Activity in Hepatocellular Carcinoma. Transl Oncol 11:1110-1118 (2018). Read more (PubMed: 30031230) »
  • Castanotto D  et al. A stress-induced response complex (SIRC) shuttles miRNAs, siRNAs, and oligonucleotides to the nucleus. Proc Natl Acad Sci U S A 115:E5756-E5765 (2018). Read more (PubMed: 29866826) »
See all 22 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (smooth muscle cells)
Specification
smooth muscle cells
Fixative
Formaldehyde
Permeabilization
Yes - 0.5% NP40/PBS
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Dr. Yiwen Liu

Verified customer

Submitted Sep 25 2012

Answer

Thank you for contacting us and for your interest in our products.

I am sorry to confirm that as far as we are aware, ab10478 has never been tested with samples from zebrafish. All tested species cross-reactivity information is stated on our datasheets, and these are updated as soon as any new information is brought to our attention.

In answer to your question, the immunogen for this antibody is as follows:

############

We recommend alignment should be over 85% to predict that an antibody will detect in a different species. If the alignment is around 80%, then theoretically there could be some crossreactivity. Please note that even if the alignment is high, we are not able to provide our guarantee for this without further testing.

If you would like to test the antibody in zebrafish, pleasedo not hesitate to contact me againby replying to this message prior to the purchase as you may be eligible for our testing discount program.

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact me for any further advice or information.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (Vascular Smooth Muscle Cell)
Loading amount
20 µg
Specification
Vascular Smooth Muscle Cell
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Andrew Cobb

Verified customer

Submitted Mar 20 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Loading amount
50 µg
Specification
HeLa
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Jan 13 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - nuclear (Mouse Embryonic Stem Cells)
Loading amount
20 µg
Specification
Mouse Embryonic Stem Cells
Gel Running Conditions
Reduced Denaturing (12% Polyacrylamide)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Apr 01 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Distal Colonic tissue Duke's Stage D Cancer)
Specification
Distal Colonic tissue Duke's Stage D Cancer
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 27°C

Abcam user community

Verified customer

Submitted Oct 27 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (CaCo 2 Cell line)
Loading amount
200 µg
Specification
CaCo 2 Cell line
Gel Running Conditions
Reduced Denaturing (15 % gel)
Blocking step
Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C

Abcam user community

Verified customer

Submitted Oct 27 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (lung carcinoma)
Specification
lung carcinoma
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH 6.0
Permeabilization
No
Blocking step
Serum as blocking agent for 10 minute(s) · Concentration: 1.5%

Abcam user community

Verified customer

Submitted Sep 26 2008

Answer

Thank you for your enquiry. We update our online datasheets with new information as soon as we receive it. As you can see from the online datasheet, on February 1st we published an Abreview from a customer who successfully used the antibody in an ICC/IF experiment. As such, this application was added to the datasheet. The online datasheet will always have the most up-to-date information. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (cervix)
Specification
cervix
Fixative
Paraformaldehyde
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2%

Dr. Alexander Rapp

Verified customer

Submitted Feb 01 2007

1-10 of 12 Abreviews or Q&A

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