Anti-CBX1 / HP1 beta antibody (ab10478)
Key features and details
- Rabbit polyclonal to CBX1 / HP1 beta
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Rat, Human
- Isotype: IgG
Overview
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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, IPmore details -
Species reactivity
Reacts with: Rat, Human
Predicted to work with: Mouse, Chicken, Hamster, Drosophila melanogaster, Marsupial
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab30861) -
Positive control
- WB: HEK293T, HAP1, MCF7, A431, HeLa and PC12 cell lysates. IP: HeLa cell lysate.
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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
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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
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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KO cell lines
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KO cell lysates
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KO cell pellets
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Recombinant Protein
Applications
The Abpromise guarantee
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 | (5) |
1/500 - 1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 21 kDa).
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| IP |
Use at an assay dependent concentration.
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| Notes |
|---|
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WB
1/500 - 1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 21 kDa). |
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IP
Use at an assay dependent concentration. |
Target
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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
modificationsNot phosphorylated.
Ubiquitinated. -
Cellular localization
Nucleus. Unassociated with chromosomes during mitosis. - Information by UniProt
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Database links
- Entrez Gene: 10951 Human
- Entrez Gene: 12412 Mouse
- Entrez Gene: 360609 Rat
- Omim: 604511 Human
- SwissProt: P83916 Human
- SwissProt: P83917 Mouse
- Unigene: 77254 Human
- Unigene: 29055 Mouse
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Alternative names
- CBX 1 antibody
- CBX antibody
- Cbx1 antibody
see all
Images
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Western blot - Anti-CBX1 / HP1 beta antibody (ab10478)All lanes : Anti-CBX1 / HP1 beta antibody (ab10478) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : CBX1 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 21 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab10478 observed at 26 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab10478 was shown to react with CBX1 / HP1 beta in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266356 (knockout cell lysate ab257383) was used. Wild-type HEK-293T and CBX1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab10478 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-CBX1 / HP1 beta antibody (ab10478)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.
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Western blot - Anti-CBX1 / HP1 beta antibody (ab10478)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 withCBX1 / HP1 beta peptide (ab30861) at 1 µg/ml
Lane 7 : HeLa nuclear lysate withCBX1 / HP1 beta peptide (ab30861) at 1 µg/ml
Lane 8 : A431 lysate withCBX1 / HP1 beta peptide (ab30861) at 1 µg/ml
Lane 9 : MCF7 cell lysate withCBX1 / HP1 beta peptide (ab30861) at 1 µg/ml
Lane 10 : HEK293 cell lysate withCBX1 / 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) -
Immunoprecipitation - Anti-CBX1 / HP1 beta antibody (ab10478)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. -
Western blot - Anti-CBX1 / HP1 beta antibody (ab10478)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 -
Western blot - Anti-CBX1 / HP1 beta antibody (ab10478)Anti-CBX1 / HP1 beta antibody (ab10478) at 1/500 dilution +Recombinant Human CBX1 / HP1 beta protein (ab109847) at 0.01 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 21 kDa
Exposure time: 10 seconds
Protocols
Datasheets and documents
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Datasheet download
References (27)
ab10478 has been referenced in 27 publications.
- Xu Y et al. The Prognostic Value of the Chromobox Family in Human Ovarian Cancer. J Cancer 11:5198-5209 (2020). PubMed: 32742466
- Gomes AP et al. Dynamic Incorporation of Histone H3 Variants into Chromatin Is Essential for Acquisition of Aggressive Traits and Metastatic Colonization. Cancer Cell 36:402-417.e13 (2019). PubMed: 31564638
- Yang YF et al. CBX1 Indicates Poor Outcomes and Exerts Oncogenic Activity in Hepatocellular Carcinoma. Transl Oncol 11:1110-1118 (2018). 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). PubMed: 29866826
- Sikorski K et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371
