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Rabbit Recombinant Monoclonal CBX4 antibody. Suitable for IP, ChIP, WB, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 3 publications.


Images

ChIP - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (AB242149), expandable thumbnail
  • Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (AB242149), expandable thumbnail
  • Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (AB242149), expandable thumbnail
  • Immunoprecipitation - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (AB242149), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (AB242149), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPChIPWBICC/IFFlow Cyt (Intra)IHC-PIHC-Fr
Human
Tested
Tested
Tested
Not recommended
Tested
Not recommended
Not recommended
Mouse
Expected
Expected
Tested
Not recommended
Expected
Tested
Not recommended
Rat
Expected
Expected
Tested
Not recommended
Expected
Tested
Not recommended

Tested
Tested

Species

Human

Dilution info

1/30

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

2 µg for 25.00000 µg chromatin

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

-

Species

Rat

Dilution info

1/1000

Notes

-

Species

Human

Dilution info

1/1000

Notes

-

Not recommended
Not recommended

Species

Human, Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/600

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/200

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Rat

Dilution info

1/200

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species

Human

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species

Rat, Mouse, Human

Dilution info

-

Notes

-

Associated Products

Select an associated product type

1 product for Alternative Product

Target data

Function

E3 SUMO-protein ligase which facilitates SUMO1 conjugation by UBE2I (PubMed:12679040). Involved in the sumoylation of HNRNPK, a p53/TP53 transcriptional coactivator, hence indirectly regulates p53/TP53 transcriptional activation resulting in p21/CDKN1A expression. Monosumoylates ZNF131 (PubMed:22825850).Component of a Polycomb group (PcG) multiprotein PRC1-like complex, a complex class required to maintain the transcriptionally repressive state of many genes, including Hox genes, throughout development (PubMed:12167701, PubMed:19636380, PubMed:21282530). PcG PRC1 complex acts via chromatin remodeling and modification of histones; it mediates monoubiquitination of histone H2A 'Lys-119', rendering chromatin heritably changed in its expressibility (PubMed:12167701, PubMed:19636380, PubMed:21282530). Binds to histone H3 trimethylated at 'Lys-9' (H3K9me3) (By similarity). Plays a role in the lineage differentiation of the germ layers in embryonic development (By similarity).

Alternative names

Recommended products

Rabbit Recombinant Monoclonal CBX4 antibody. Suitable for IP, ChIP, WB, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 3 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR23053-7

Purification technique

Affinity purification Protein A

Specificity

The human recommendation is based on the WB results. We do not guarantee IHC-P for human.

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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 monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

CBX4 also known as Chromobox protein homolog 4 or Polycomb 2 acts mechanically as part of the Polycomb group (PcG) proteins. It weighs approximately 60 kDa and it is found in nuclear matrix where it participates in chromatin remodeling. CBX4 shows significant expression in tissues like liver brain and lung indicating its wide importance to cellular function. By binding to methylated histones especially H3K27me3 CBX4 influences gene silencing and epigenetic regulation important for maintaining cellular identity.

Biological function summary

CBX4 operates as a component of the Polycomb Repressive Complex 1 (PRC1) which plays role in controlling gene expression patterns during development. In this complex CBX4 interacts with other core components such as BMI1 and RING1 to facilitate monoubiquitination of histone H2A leading to transcriptional repression. This ability of CBX4 to modulate epigenetic states highlights its importance in critical developmental processes and differentiation.

Pathways

CBX4 significantly influences gene regulatory networks involved in cell cycle and apoptosis pathways. In the cell cycle regulation CBX4 interacts with important proteins such as Cyclin D1 impacting cell proliferation. In apoptosis CBX4 has roles linked to the p53 pathway by modulating transcriptional activity which influences cellular fate. These pathways underline the significance of CBX4 in maintaining balance between cell growth and death.

Associated diseases and disorders

The deregulation of CBX4 expression has been associated with cancers like hepatocellular carcinoma and breast cancer. CBX4 potentially interacts with proteins such as p21 and p16 whose pathways play significant roles in cancer development. This underlines its role as a potential biomarker or therapeutic target in cancer treatment strategies. Understanding the function and regulation of CBX4 could offer insights in devising novel treatments for these malignancies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

  • ChIP - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149), expandable thumbnail

    ChIP - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149)

    Chromatin was prepared from human NCCIT cells according to the Abcam Dual-X-ChIP protocol. Cells were fixed with 1.5 mM EGS for 30mins and then 1% formaldehyde for 10min.

    The ChIP was performed with 25 μg of chromatin, 2 μg of ab242149 (red), or 2 μg of rabbit normal IgG Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 (gray) and 20 μl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).

  • Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149), expandable thumbnail

    Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149)

    False colour image of Western blot: Anti-CBX4 antibody [EPR23053-7] - ChIP Grade staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab242149 was shown to bind specifically to CBX4. A band was observed at 75 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in Cbx4 knockout cell line Human CBX4 knockout HEK-293 cell line ab261723 (knockout cell lysate ab261666). The band observed in the knockout lysate lane below 75 kDa is likely to represent a truncated form of CBX4. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and Cbx4 knockout HEK-293 cell lysates were analysed.First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

    All lanes: Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149) at 1/1000 dilution

    Lane 1: Wild-type HEK-293 cell lysate at 20 µg

    Lane 2: Cbx4 knockout HEK-293 cell lysate at 20 µg

    Lane 3: RAW 264.7 cell lysate at 20 µg

    Lane 4: HUVEC cell lysate at 20 µg

    Performed under reducing conditions.

  • Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149), expandable thumbnail

    Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149)

    False colour image of Western blot: Anti-CBX4 antibody [EPR23053-7] - ChIP Grade staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab242149 was shown to bind specifically to CBX4. A band was observed at 75 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in Cbx4 CRISPR-Cas9 edited cell line Human CBX4 knockout HEK-293 cell line ab261723 (CRISPR-Cas9 edited cell lysate ab261666). The band observed in the CRISPR-Cas9 edited lysate lane below 75 kDa is likely to represent a truncated form of CBX4. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and Cbx4 CRISPR-Cas9 edited HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

    All lanes: Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149) at 1/1000 dilution

    Lane 1: Wild-type HEK-293 cell lysate at 20 µg

    Lane 2: Cbx4 CRISPR-Cas9 edited HEK-293 cell lysate at 20 µg

    Lane 3: RAW 264.7 cell lysate at 20 µg

    Lane 4: HUVEC cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 61 kDa

    Observed band size: 75 kDa

  • Immunoprecipitation - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149), expandable thumbnail

    Immunoprecipitation - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149)

    CBX4 was immunoprecipitated from 0.35 mg HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate with ab242149 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab242149 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

    Lane 1: HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate 10ug

    Lane 2: ab242149 IP in HCT 116 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab242149 in HCT 116 whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 15 seconds.

    All lanes: Immunoprecipitation - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149)

    Predicted band size: 52 kDa, 61 kDa

    Observed band size: 45 kDa, 60 kDa, 66 kDa, 74 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149)

    Immunohistochemical analysis of paraffin-embedded rat hippocampus tissue labeling CBX4 with ab242149 at 1/200 dilution (2.785 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Nuclear staining on rat hippocampus (PMID: 28283560). The section was incubated with ab242149 for 30 at RT. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

  • Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149), expandable thumbnail

    Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149)

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 26 seconds.

    The molecular weight observed is consistent with what has been described in the literature (PMID: 27864346).

    All lanes: Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149) at 1/1000 dilution

    Lane 1: HCT 116 (human colorectal carcinoma epithelial cell), whole cell lysate at 20 µg

    Lane 2: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 20 µg

    Lane 3: C6 (rat glial tumor glial cell), whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 61 kDa

    Observed band size: 74 kDa

  • Flow Cytometry (Intracellular) - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149)

    Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NCCIT (Human pluripotent embryonic carcinoma epithelial cell) cells labelling CBX4 with ab242149 at 1/600 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149)

    Immunohistochemical analysis of paraffin-embedded Mouse hippocampus tissue labeling CBX4 with ab242149 at 1/200 dilution (2.785 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Nuclear staining on mouse hippocampus (PMID: 28283560). The section was incubated with ab242149 for 30 at RT. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149)

    Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling CBX4 with ab242149 at 1/200 dilution (2.785 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Nuclear staining on mouse cerebrum (PMID: 28283560). The section was incubated with ab242149 for 30 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

  • Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149), expandable thumbnail

    Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149)

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

    The molecular weight observed is consistent with what has been described in the literature (PMID: 27864346).

    All lanes: Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149) at 1/1000 dilution

    Lane 1: HDLM-2 (human Hodgkin lymphoma) whole cell lysate at 20 µg

    Lane 2: NCCIT (human pluripotent embryonic carcinoma epithelial cell), whole cell lysate at 20 µg

    Lane 3: Mouse thymus tissue lysate at 20 µg

    Lane 4: Rat thymus tissue lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 61 kDa

    Observed band size: 74 kDa

  • Flow Cytometry (Intracellular) - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149)

    Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HCT 116 (Human colorectal carcinoma epithelial cell) cells labelling CBX4 with ab242149 at 1/600 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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